Novel egfr modulators and uses thereof

ABSTRACT

The present invention relates to certain pyrrolopyrimidine derivatives, pharmaceutical compositions containing them, and methods of using them, including methods for the treatment of tumors and related diseases related to the dysregulation of kinase (such as EGFR (including HER), Alk, PDGFR, but not limited to) pathways.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application, in certain aspects, relates to U.S. ProvisionalApplication No. 61/586,718, filed Jan. 13, 2012, the disclosure of whichis hereby incorporated by reference in its entirety. This applicationclaims priority to U.S. Provisional Application No. 61/680,231, filedAug. 6, 2012, the disclosure of which is hereby incorporated byreference in its entirety.

TECHNICAL FIELD

The field of this invention is pharmaceutical compounds, compositionsand methods, especially as they are related to compositions and methodsfor the treatment of proliferation disorders and related diseasesrelated to the dysregulation of kinase (such as, but not limited to,EGFR (including HER), Alk, and PDGFR) pathways.

BACKGROUND ART

Protein kinases are a group of enzymes that regulate diverse, importantbiological processes including cell growth, proliferation, survival,invasion and differentiation, organ formation, tissue repair andregeneration, etc. Protein kinases exert their physiological functionsthrough catalyzing the phosphorylation of protein and thereby modulatingthe cellular activities. Because protein kinases have profound effectson cells, their activities are highly regulated. Kinases are turned onor off by phosphorylation (sometimes by autophosphorylation), by bindingof activator proteins or inhibitor proteins, or small molecules, or bycontrolling their location in the cell relative to their substrates.Dysfunctions in the activities of kinases, arising from geneticabnormalities or environmental factors, are known to be associated withmany diseases. Several severe pathological states, including cancer andchronic inflammation, are associated with stimulation of intra-cellularsignaling, and since kinases positively relay signaling events, theirinhibition offers a powerful way to inhibit or control signaltransduction cascades.

The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is amember of the ErbB family of receptors, a subfamily of four closelyrelated receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2),Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR is the cell-surface receptor formembers of the epidermal growth factor family (EGF-family) ofextracellular protein ligands. Mutations affecting EGFR expression oractivity could result in cancer. EGFR is reported deregulated in mostsolid tumor types i.e. lung cancer, breast cancer and brain tumor. It isestimated that mutations, amplifications or misregulations of EGFR orfamily members are implicated in about 30% of all epithelial cancers.Therapeutic approaches have been developed based on the inhibition ofEGFR by either antibody drug or small molecular inhibitor drug, such asgefitinib and erlotinib. In the case of non-small cell lung cancer,gefitinib and erlotinib have shown benefit for about 10-40% of thepatients. However, acquired resistant to gefitinib or erlotinib after aperiod of treatment become a major clinical problem. Research hasconfirmed that one main reason resistance developed is due to thepresence of a new mutation of T790M, which is the gatekeeper of EGFR.Subsequently, inhibitors can overcome this T790M have been developed andshowed advantage in the clinical trial, such as BIBW2992. However, theseT790M targeted EGFR inhibitor still has relative inhibitory activitytowards wild type EGFR which limit the clinical application. It isneeded to further develop more efficient type of EGFR inhibitor whichwill target substantially the mutation and not substantially the wildtype protein.

SUMMARY

The present invention is directed to certain pyrrolopyrimidinederivatives, pharmaceutical compositions, and methods of using thesecompounds and compositions to treat proliferation disorders.

The present disclosure provides a compound of Formula (VIII):

wherein

X¹ is O, NH, S, CH₂, or CF₂;

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl;

R¹³ is selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl, SO₂—C₁₋₆alkyl,C₃₋₇ cycloalkyl, and C₆₋₂₀ aryl,

-   -   wherein each alkyl or aryl is unsubstituted or substituted with        hydroxyl, C₁₋₆ alkoxy, or halo; and

—NR¹⁸R¹⁹ is

-   -   wherein R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;    -   R¹⁵ is unsubstituted methyl, or is C₂₋₄alkyl unsubstituted or        substituted with hydroxy, methoxy, or halo; and    -   m is 1 or 2;

or R¹⁹ and R⁹ taken together form a 5- or 6-membered heteroaryl ringoptionally substituted with C₁₋₆alkyl that is unsubstituted orsubstituted with amino, hydroxyl, or halo; and R¹⁸ is hydrogen orC₁₋₆alkyl, or is absent to satisfy valency of the heteroaryl ring;

provided that neither of R⁶ or R⁷ is methoxy when —NR¹⁸R¹⁹ is

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (Ia) and (Ib):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

-   -   a is one or two;

Ring A is an aromatic ring;

R²⁰ and R²¹ are independently selected from hydrogen and C₁₋₆ alkyl;wherein alkyl is unsubstituted or substituted with amino, hydroxyl, orhalo; wherein R²¹ may not present to satisfy valency;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl, SO₂—C₁₋₆alkyl,C₃₋₇ cycloalkyl, and C₆₋₂₀ aryl,

-   -   wherein each alkyl or aryl is unsubstituted or substituted with        hydroxyl, C₁₋₆ alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (II):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (III):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and R¹³ is selected fromhydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein each alkyl or aryl isunsubstituted or substituted with hydroxyl, C₁₋₆ alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (IV):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (V):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

The present disclosure provides a compound of Formula (VI):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In certain embodiments, the present disclosure provides a compound ofFormula (VII) as described below.

In certain embodiments, the compound of Formula (I)-(VIII) is a compoundselected from those species described or exemplified in the detaileddescription below.

In a further aspect, the present disclosure provides a pharmaceuticalcomposition comprising at least one compound of Formula (I)-(VIII) or apharmaceutically acceptable salt thereof. Pharmaceutical compositionsmay further comprise a pharmaceutically acceptable carrier or excipient.The present disclosure also provides a compound of Formula (I)-(VIII) ora pharmaceutically acceptable salt thereof for use as a medicament.

In another aspect, the present disclosure provides a method of treatingand/or preventing a proliferation disorder comprising administering to asubject in need of such treatment an effective amount of at least onecompound of Formula (I)-(VIII) or a pharmaceutically acceptable saltthereof.

In another aspect, the present disclosure provides a compound of Formula(I)-(VIII) or a pharmaceutically acceptable salt thereof for use intherapy. In another aspect, the present disclosure provides a compoundof Formula (I)-(VIII) or a pharmaceutically acceptable salt thereof foruse in the treatment of a proliferation disorder. In another aspect, thepresent disclosure provides use of a compound of Formula (I)-(VIII) or apharmaceutically acceptable salt thereof for the manufacture of amedicament for the treatment of a proliferation disorder.

In another aspect, the present disclosure provides a method of treatinga condition associated with EGFR inhibitory activity targeting a mutatedEGFR but not the wild type EGFR comprising administering to a subject inneed of such treatment an effective amount of at least one compound ofFormula (I)-(VIII) or a pharmaceutically acceptable salt thereof. Insome embodiments, the mutated EGFR comprises a T790M mutation. Thepresent disclosure provides use of a compound of Formula (I)-(VIII) inthe preparation of a medicament for the treatment of such diseases andmedical conditions, and the use of such compounds and salts fortreatment of such diseases and medical conditions.

In another aspect, the present disclosure provides a method ofinhibiting mutated EGFR in a cell comprising contacting the cell with aneffective amount of at least one compound of Formula (I)-(VIII) or asalt thereof, and/or with at least one pharmaceutical composition of theembodiments, wherein the contacting is in vitro, ex vivo, or in vivo. Insome embodiments, the mutated EGFR comprises a T790M mutation.

One of ordinary skill in the art will recognize that compounds ofFormula (II)-(VI) are compounds of Formula (I), and that compounds ofFormula (I)-(VII) are compounds of Formula (VIII).

Additional embodiments, features, and advantages of the invention willbe apparent from the following detailed description and through practiceof the invention.

For the sake of brevity, the disclosures of the publications cited inthis specification, including patents, are herein incorporated byreference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows SDS-PAGE of certain effectors of H1975 lung cancer cellstreated with varying concentrations of Compound 3.

FIG. 2 shows immunoblots of certain effectors in tumors treated withCompound 3 at various time intervals.

FIG. 3 shows a chart of mouse body weight changes in the differentgroups in NCI-H1975 model.

FIG. 4 shows a chart of mouse body weight changes in the differentgroups in HCC827 model.

FIG. 5 shows a chart of mouse body weight changes in the differentgroups in A431 model.

FIG. 6 shows a chart of the tumor volume of mice in the different groupsin NCI-H1975 model.

FIG. 7 shows a chart of the tumor volume of mice in the different groupsin HCC827 model.

FIG. 8 shows a chart of the tumor volume of mice in the different groupsin A431 model.

FIGS. 9A-9F show SDS-PAGE (9A, 9C, 9E) and inhibition graphs (9B, 9D,9F) of EGFR-Tyr1068 phosphorylation and downstream signaling in H1975lung cancer cells treated with varying concentrations of Compound 3 (9A,9B), Gefitinib (9C, 9D), and WZ4002 (9E, 9F).

FIGS. 10A-10F show SDS-PAGE (10A, 10C, 10E) and inhibition graphs (10B,10D, 10F) of EGFR-Tyr1068 phosphorylation and downstream signaling inHCC-827 EGFR mutant cells treated with varying concentrations ofCompound 3 (10A, 10B), Gefitinib (10C, 10D), and WZ4002 (10E, 10F).

FIGS. 11A-11F show SDS-PAGE (11A, 11C, 11E) and inhibition graphs (11B,11D, 11F) of EGFR-Tyr1068 phosphorylation and downstream signaling inA431 cells expressing WT EGFR that were treated with varyingconcentrations of Compound 3 (11A, 11B), Gefitinib (11C, 11D), andWZ4002 (11E, 11F).

FIG. 12 shows the inhibition of phosphorylation of EGFR in H1975 tumortissues when treated with a single dose of Compound 3 at 12.5, 50, and200 mg/kg.

FIG. 13 shows the inhibition of phosphorylation of EGFR in H1975 tumortissues when treated with eight doses of Compound 3 at 12.5 and 50mg/kg, as compared to Gefitinib at 100 mg/kg.

FIG. 14 shows the results of a cell-based pulse-chase assay thatdemonstrate that Compound 3 is an irreversible inhibitor ofproliferation of H1975 cells with the EGFR T790M mutation.

DETAILED DESCRIPTION

The present invention is directed to certain pyrrolopyrimidinederivatives, pharmaceutical compositions, and methods of using thesecompounds and compositions to treat proliferation disorders. Thecompounds as described herein exhibit anti-tumor, anticancer,anti-inflammation, anti-infectious, and anti-proliferation activity. Insome embodiments, the compounds have been shown to possess anti-canceractivity in cell based assays as described herein using various cancercell lines, which demonstrate very efficient EGFR inhibitory activitytargeting substantially the mutation and not substantially the wild typeprotein. In some embodiments, the mutated EGFR comprises a T790Mmutation. Accordingly, the compounds and compositions comprising thecompounds of the embodiments are useful to treat conditionscharacterized by those mutated cancer cells. In certain instances, thecompounds are useful to treat sarcoma, epidermoid cancer, fibrosarcoma,cervical cancer, gastric carcinoma, skin cancer, leukemia, lymphoma,lung cancer, non-small cell lung cancer, colon cancer, CNS cancer,melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer,liver cancer, head and neck cancers, and pancreatic cancer.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

As used herein, the terms “including,” “containing,” and “comprising”are used in their open, non-limiting sense.

To provide a more concise description, some of the quantitativeexpressions given herein are not qualified with the term “about”. It isunderstood that, whether the term “about” is used explicitly or not,every quantity given herein is meant to refer to the actual given value,and it is also meant to refer to the approximation to such given valuethat would reasonably be inferred based on the ordinary skill in theart, including equivalents and approximations due to the experimentaland/or measurement conditions for such given value. Whenever a yield isgiven as a percentage, such yield refers to a mass of the entity forwhich the yield is given with respect to the maximum amount of the sameentity that could be obtained under the particular stoichiometricconditions. Concentrations that are given as percentages refer to massratios, unless indicated differently.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

Except as otherwise noted, the methods and techniques of the presentembodiments are generally performed according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout the presentspecification. See, e.g., Loudon, Organic Chemistry, Fourth Edition, NewYork: Oxford University Press, 2002; Smith and March, March's AdvancedOrganic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition,Wiley-Interscience, 2001.

The nomenclature used herein to name the subject compounds isillustrated in the Examples herein. This nomenclature has generally beenderived using the commercially-available AutoNom software (MDL, SanLeandro, Calif.).

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. All combinations of the embodimentspertaining to the chemical groups represented by the variables arespecifically embraced by the present invention and are disclosed hereinjust as if each and every combination was individually and explicitlydisclosed, to the extent that such combinations embrace compounds thatare stable compounds (i.e., compounds that can be isolated,characterized, and tested for biological activity). In addition, allsubcombinations of the chemical groups listed in the embodimentsdescribing such variables are also specifically embraced by the presentinvention and are disclosed herein just as if each and every suchsub-combination of chemical groups was individually and explicitlydisclosed herein.

Chemical Terms

The term “alkyl” refers to a straight- or branched-chain alkyl grouphaving from 1 to 12 carbon atoms in the chain. Examples of alkyl groupsinclude methyl (Me), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl,sec-butyl, tert-butyl (tBu), pentyl, isopentyl, tert-pentyl, hexyl,isohexyl, and groups that in light of the ordinary skill in the art andthe teachings provided herein would be considered equivalent to any oneof the foregoing examples.

The term “alkoxy” refers to an alkyl group as defined above, bonded toan oxygen atom. The alkoxy group is connected to the parent structurevia the oxygen atom.

The term “amino” refers to an —NH₂ group, or a mono- or dialkylaminogroup.

The term “cycloalkyl” refers to a saturated or partially saturated,monocyclic, fused polycyclic, bridged polycyclic, or spiro polycycliccarbocycle having from 3 to 12 ring atoms per carbocycle. Illustrativeexamples of cycloalkyl groups include the following entities, in theform of properly bonded moieties:

The term “heteroaryl” refers to a monocyclic, fused bicyclic, or fusedpolycyclic aromatic heterocycle (ring structure having ring atomsselected from carbon atoms and up to four heteroatoms selected fromnitrogen, oxygen, and sulfur) having from 3 to 12 ring atoms perheterocycle. Illustrative examples of heteroaryl groups include thefollowing entities, in the form of properly bonded moieties:

The term “halogen” represents chlorine, fluorine, bromine, or iodine.The term “halo” represents chloro, fluoro, bromo, or iodo. The term“haloalkyl” means an alkyl as defined above, substituted with one ormore halogen atoms. The term “haloalkoxy” means an alkoxy as definedabove, substituted with one or more halogen atoms.

The term “acyl” refers to a group R—C(O)— of from 1 to 10 carbon atomsof a straight, branched, or cyclic configuration or a combinationthereof, attached to the parent structure through carbonylfunctionality. Such group may be saturated or unsaturated, and aliphaticor aromatic.

The term “cyano” refers to the group —CN.

The term “nitro” refers to the group —NO₂.

The term “hydroxyl” refers to the group —OH.

Those skilled in the art will recognize that the species listed orillustrated above are not exhaustive, and that additional species withinthe scope of these defined terms may also be selected.

The term “substituted” means that the specified group or moiety bearsone or more substituents. The term “unsubstituted” means that thespecified group bears no substituents. The term “optionally substituted”means that the specified group is unsubstituted or substituted by one ormore substituents. Where the term “substituted” is used to describe astructural system, the substitution is meant to occur at anyvalency-allowed position on the system.

Any formula depicted herein is intended to represent a compound of thatstructural formula as well as certain variations or forms. For example,a formula given herein is intended to include a racemic form, or one ormore enantiomeric, diastereomeric, or geometric isomers, or a mixturethereof. Additionally, any formula given herein is intended to referalso to a hydrate, solvate, or polymorph of such a compound, or amixture thereof.

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the embodiments include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, andiodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S,¹⁸F, ³⁶Cl, and ¹²⁵I, respectively. Such isotopically-labelled compoundsare useful in metabolic studies (preferably with ¹⁴C), reaction kineticstudies (with, for example ²H or ³H), detection or imaging techniques[such as positron emission tomography (PET) or single-photon emissioncomputed tomography (SPECT)] including drug or substrate tissuedistribution assays, or in radioactive treatment of patients. Inparticular, an ¹⁸F or ¹¹C labeled compound may be particularly preferredfor PET or SPECT studies. Further, substitution with heavier isotopessuch as deuterium (i.e., ²H) may afford certain therapeutic advantagesresulting from greater metabolic stability, for example increased invivo half-life or reduced dosage requirements. Isotopically-labeledcompounds of the embodiments and prodrugs thereof can generally beprepared by carrying out the procedures disclosed in the schemes or inthe examples and preparations described below by substituting a readilyavailable isotopically-labeled reagent for a non-isotopically-labeledreagent.

The nomenclature “C_(i-j)” with j>i, when applied herein to a class ofsubstituents, is meant to refer to embodiments for which each and everyone of the number of carbon members, from i to j including i and j, isindependently realized. By way of example, the term C₁₋₃ refersindependently to embodiments that have one carbon member (C₁),embodiments that have two carbon members (C₂), and embodiments that havethree carbon members (C₃).

Any disubstituent referred to herein is meant to encompass the variousattachment possibilities when more than one of such possibilities areallowed. For example, reference to disubstituent -A-B—, where A≠B,refers herein to such disubstituent with A attached to a firstsubstituted member and B attached to a second substituted member, and italso refers to such disubstituent with A attached to the secondsubstituted member and B attached to the first substituted member.

The present disclosure provides pharmaceutically acceptable salts of thecompounds represented by Formulae (I)-(VIII), preferably of thosedescribed above and of the specific compounds exemplified herein, andpharmaceutical compositions comprising such salts, and methods of usingsuch salts.

A “pharmaceutically acceptable salt” is intended to mean a salt of afree acid or base of a compound represented herein that is non-toxic,biologically tolerable, or otherwise biologically suitable foradministration to the subject. See, generally, S. M. Berge, et al.,“Pharmaceutical Salts,” J. Pharm. Sci., 1977, 66, 1-19. Preferredpharmaceutically acceptable salts are those that are pharmacologicallyeffective and suitable for contact with the tissues of subjects withoutundue toxicity, irritation, or allergic response. A compound describedherein may possess a sufficiently acidic group, a sufficiently basicgroup, both types of functional groups, or more than one of each type,and accordingly react with a number of inorganic or organic bases, andinorganic and organic acids, to form a pharmaceutically acceptable salt.

Examples of pharmaceutically acceptable salts include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogen-phosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,methylsulfonates, propylsulfonates, besylates, xylenesulfonates,naphthalene-1-sulfonates, naphthalene-2-sulfonates, phenylacetates,phenylpropionates, phenylbutyrates, citrates, lactates,γ-hydroxybutyrates, glycolates, tartrates, and mandelates.

For a compound of Formulae (I)-(VIII) that contains a basic nitrogen, apharmaceutically acceptable salt may be prepared by any suitable methodavailable in the art, for example, treatment of the free base with aninorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuricacid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and thelike, or with an organic acid, such as acetic acid, phenylacetic acid,propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid,hydroxymaleic acid, isethionic acid, succinic acid, valeric acid,fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid,salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidylacid, such as glucuronic acid or galacturonic acid, an alpha-hydroxyacid, such as mandelic acid, citric acid, or tartaric acid, an aminoacid, such as aspartic acid or glutamic acid, an aromatic acid, such asbenzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, asulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid,methanesulfonic acid, or ethanesulfonic acid, or any compatible mixtureof acids such as those given as examples herein, and any other acid andmixture thereof that are regarded as equivalents or acceptablesubstitutes in light of the ordinary level of skill in this technology.In certain embodiments, the pharmaceutically acceptable salt is the HClsalt, maleic acid salt, HBr salt, lactic acid salt, tartaric acid salt,or methanesulfonic acid salt.

The present disclosure provides pharmaceutically acceptable prodrugs ofthe compounds of Formulae (I)-(VIII), and treatment methods employingsuch pharmaceutically acceptable prodrugs. The term “prodrug” means aprecursor of a designated compound that, following administration to asubject, yields the compound in vivo via a chemical or physiologicalprocess such as solvolysis or enzymatic cleavage, or under physiologicalconditions (e.g., a prodrug on being brought to physiological pH isconverted to the compound of Formulae (I)-(VIII)). A “pharmaceuticallyacceptable prodrug” is a prodrug that is non-toxic, biologicallytolerable, and otherwise biologically suitable for administration to thesubject. Illustrative procedures for the selection and preparation ofsuitable prodrug derivatives are described, for example, in “Design ofProdrugs”, ed. H. Bundgaard, Elsevier, 1985.

The present disclosure provides pharmaceutically active metabolites ofcompounds of Formulae (I)-(VIII), and uses of such metabolites in themethods of the embodiments. A “pharmaceutically active metabolite” meansa pharmacologically active product of metabolism in the body of acompound of Formulae (I)-(VIII) or salt thereof. Prodrugs and activemetabolites of a compound may be determined using routine techniquesknown or available in the art. See, e.g., Bertolini et al., J. Med.Chem. 1997, 40, 2011-2016; Shan et al., J. Pharm. Sci. 1997, 86 (7),765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230; Bodor, Adv. DrugRes. 1984, 13, 255-331; Bundgaard, Design of Prodrugs (Elsevier Press,1985); and Larsen, Design and Application of Prodrugs, Drug Design andDevelopment (Krogsgaard-Larsen et al., eds., Harwood AcademicPublishers, 1991).

Representative Embodiments Formula (VIII)

The present disclosure provides a compound of Formula (VIII). In someembodiments, X¹ is O or NH. In other embodiments, X¹ is CH₂ or CF₂. Instill other embodiments, X¹ is O.

In some embodiments of Formula (VIII), —NR¹⁸R¹⁹ is

In other embodiments, —NR¹⁸R¹⁹ is

In some embodiments, R¹⁵ is methyl, hydroxyethyl, methoxyethyl, orfluoroethyl. In other embodiments, R¹⁵ is fluoroethyl. In someembodiments, m is 1. In other embodiments, m is 2.

In some embodiments, R⁹ and R¹⁹ taken together form an optionallysubstituted 5- or 6-membered heteroaryl ring. In some embodiments, R¹⁹and R⁹ together form a 5- or 6-membered ring optionally substituted withC₁₋₆alkyl that is unsubstituted or substituted with amino. In someembodiments, the heteroaryl ring is substituted with dimethylaminomethylor piperidinylmethyl. In other embodiments, R⁹ and R¹⁹ taken togetherform pyrrole or pyridine. In some embodiments, R¹⁸ isdimethylaminoethyl.

In some embodiments, R⁶ is methoxy. In other embodiments, R⁷ is methoxy.In certain instances, R⁷ is hydrogen or methoxy.

In some embodiments of Formula (VIII), each variable therein is definedas described below for any one of Formulas (I)-(VII) or embodimentsthereof. In particular, certain embodiments of Formula (VIII) are asdefined for each variable for Formula (I) below, and said definitionsare incorporated herein by reference.

Formula (I)

The present disclosure provides a compound of Formula (Ia) and (Ib):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted withhydroxyl or amino; and

wherein each R²² and R²³ are independently selected from hydrogen andC₁₋₆ alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

a is one or two;

Ring A is an aromatic ring;

R²⁰ and R²¹ are independently selected from hydrogen and C₁₋₆ alkyl;wherein alkyl is unsubstituted or substituted with amino, hydroxyl, orhalo; wherein R²¹ may not present to satisfy valency;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl, SO₂—C₁₋₆alkyl,C₃₋₇ cycloalkyl, and C₆₋₂₀ aryl,

wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

Formula (I) is meant to refer to Formula (Ia) and Formula (Ib).

In Formula (I), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are each hydrogen.

In Formula (I), n is a number from zero to 4. In certain instances, n iszero. In certain instances, n is one. In certain instances, n is 2. Incertain instances, n is 3. In certain instances, n is 4.

In Formula (I), R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆alkoxy, cyano, and nitro. In certain instances, R³ is halo. In certaininstances, R³ is hydroxyl. In certain instances, R³ is C₁₋₆ alkyl. Incertain instances, R³ is C₁₋₆ alkoxy. In certain instances, R³ is cyano.In certain instances, R³ is nitro.

In Formula (I), R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇cycloalkyl, and —NR²²R²³; wherein the alkyl or cycloalkyl isunsubstituted or substituted with hydroxyl or amino; and wherein eachR²² and R²³ are independently selected from hydrogen and C₁₋₆ alkyl orR²² and R²¹ may be joined to form a 3 to 10 membered ring. In certaininstances, R⁴ is hydrogen. In certain instances, R⁴ is C₁₋₆ alkyl. Incertain instances, R⁴ is C₃₋₇ cycloalkyl. In certain instances, R⁴ is—NR²²R²³.

In certain instances, R⁴ is unsubstituted C₁₋₆ alkyl. In certaininstances, R⁴ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with hydroxyl.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with amino.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₆ alkyl that is substituted with —N(CH₃)₂.In certain instances, R⁴ is C₁₋₃ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with —N(CH₃)₂.

In certain instances, R⁴ is unsubstituted C₃₋₇ cycloalkyl. In certaininstances, R⁴ is unsubstituted C₃ cycloalkyl. In certain instances, R⁴is unsubstituted C₄ cycloalkyl. In certain instances, R⁴ isunsubstituted C₅₋₆ cycloalkyl. In certain instances, R⁴ is unsubstitutedC₇ cycloalkyl.

In certain instances, R⁴ is —NR²²R²³, wherein each R²² and R²³ areindependently selected from hydrogen and C₁₋₆ alkyl or R²² and R²³ maybe joined to form a 3 to 10 membered ring. In certain instances, R²² andR²³ are hydrogen. In certain instances, R²² and R²³ are C₁₋₆ alkyl. Incertain instances, R²² and R²³ are C₁₋₃ alkyl. In certain instances, R²²and R²³ are methyl.

In certain instances, R²² and R²³ may be joined to form a 3 to 10membered ring, such that R⁴ is

where w is a number from 1 to 8. In certain instances, R²² and R²³ maybe joined to form a 3-membered ring. In certain instances, R²² and R²³may be joined to form a 4-membered ring. In certain instances, R²² andR²³ may be joined to form a 5-membered ring. In certain instances, R²²and R²³ may be joined to form a 6-membered ring. In certain instances,R²² and R²³ may be joined to form a 7-membered ring.

In Formula (I), R⁵ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R⁵ is hydrogen. In certain instances, R⁵ is C₁₋₆ alkyl. Incertain instances, R⁵ is methyl. In certain instances, R⁵ is ethyl. Incertain instances, R⁵ is C₁₋₃ alkyl. In certain instances, R⁵ is C₄₋₆alkyl.

In Formula (I), R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁶ is hydrogen. In certain instances, R⁶ is halo. Incertain instances, R⁶ is fluoro. In certain instances, R⁶ is chloro. Incertain instances, R⁶ is bromo. In certain instances, R⁶ is C₁₋₆ alkyl.In certain instances, R⁶ is C₁₋₆ haloalkyl. In certain instances, R⁶ isC₂₋₆ alkoxy. In certain instances, R⁶ is C₁₋₆ haloalkoxy. In certaininstances, R⁶ is hydroxyl. In certain instances, R⁶ is cyano. In certaininstances, R⁶ is nitro.

In Formula (I), R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁷ is hydrogen. In certain instances, R⁷ is halo. Incertain instances, R⁷ is fluoro. In certain instances, R⁷ is chloro. Incertain instances, R⁷ is bromo. In certain instances, R⁷ is C₁₋₆ alkyl.In certain instances, R⁷ is C₁₋₆ haloalkyl. In certain instances, R⁷ isC₂₋₆ alkoxy. In certain instances, R⁷ is C₁₋₆ haloalkoxy. In certaininstances, R⁷ is hydroxyl. In certain instances, R⁷ is cyano. In certaininstances, R⁷ is nitro.

In Formula (I), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (I), Q is CR⁹ or N. In certain instances, Q is CR⁹. Incertain instances, Q is N.

In Formula (I), R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁹ is hydrogen. In certain instances, R⁹ is halo. Incertain instances, R⁹ is fluoro. In certain instances, R⁹ is chloro. Incertain instances, R⁹ is bromo. In certain instances, R⁹ is C₁₋₆ alkyl.In certain instances, R⁹ is C₁₋₆ haloalkyl. In certain instances, R⁹ isC₁₋₆ alkoxy. In certain instances, R⁹ is C₁₋₆ haloalkoxy. In certaininstances, R⁹ is hydroxyl. In certain instances, R⁹ is cyano. In certaininstances, R⁹ is nitro. In certain instances, R⁹ is hydrogen or fluoro.

In Formula (I), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆ alkyl. Incertain instances, R¹⁰ is methyl. In certain instances, R¹⁰ is ethyl. Incertain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances, R¹⁰ is C₄₋₆alkyl.

In Formula (I), a is one or two and Ring A is an aromatic ring. Incertain instances, a is one, as shown:

In certain instances, a is two, as shown:

In Formula (I), R²⁰ and R²¹ are independently selected from hydrogen andC₁₋₆ alkyl; wherein alkyl is unsubstituted or substituted with amino,hydroxyl, or halo; wherein R²¹ may not present to satisfy valency. Incertain instances, R²⁰ and R²¹ are independently hydrogen. In certaininstances, R²⁰ and R²¹ are independently unsubstituted C₁₋₆ alkyl. Incertain instances, R²⁰ and R²¹ are independently C₁₋₆ alkyl, substitutedwith amino. In certain instances, R²⁰ and R²¹ are independently C₁₋₆alkyl, substituted with —NR²⁴R²⁵, wherein R²⁴ and R²⁵ are independentlyselected from hydrogen and C₁₋₆alkyl. In certain instances, R²⁰ and R²¹are independently C₁₋₆ alkyl, substituted with —NR²⁴R²⁵, wherein R²⁴ andR²⁵ are independently selected from hydrogen and C₁₋₃alkyl. In certaininstances, R²⁰ and R²¹ are independently C₁₋₆ alkyl, substituted with—NR²⁴R²⁵, wherein R²⁴ and R²⁵ are independently selected from hydrogenand methyl. In certain instances, R²⁰ and R²¹ are independently C₁₋₃alkyl, substituted with —NR²⁴R²⁵, wherein R²⁴ and R²⁵ are independentlyselected from hydrogen and methyl. In certain instances, R²⁰ and R²¹ areindependently C₁₋₆ alkyl, substituted with hydroxyl. In certaininstances, R²⁰ and R²¹ are C₁₋₆ alkyl, substituted with halo.

In Formula (I), R¹¹ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹¹ is hydrogen. In certain instances, R¹¹ is C₁₋₆ alkyl. Incertain instances, R¹¹ is methyl. In certain instances, R¹¹ is ethyl. Incertain instances, R¹¹ is C₁₋₃ alkyl. In certain instances, R¹¹ is C₄₋₆alkyl.

In Formula (I), R¹² is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹² is hydrogen. In certain instances, R¹² is C₁₋₆ alkyl. Incertain instances, R¹² is methyl. In certain instances, R¹² is ethyl. Incertain instances, R¹² is C₁₋₃ alkyl. In certain instances, R¹² is C₄₋₆alkyl.

In Formula (I), R¹³ is selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl,SO₂—C₁₋₆alkyl, C₃₋₇ cycloalkyl, and C₆₋₂₀ aryl, wherein each alkyl oraryl is unsubstituted or substituted with hydroxyl, C₁₋₆ alkoxy, orhalo. In certain instances, R¹³ is hydrogen. In certain instances, R¹³is C₁₋₆ alkyl. In certain instances, R¹³ is C₁₋₆ acyl. In certaininstances, R¹³ is SO₂—C₁₋₆alkyl. In certain instances, R¹³ is C₃₋₇cycloalkyl. In certain instances, R¹³ is C₆₋₂₀ aryl. In certaininstances, R¹³ is C₁₋₆alkyl substituted with hydroxyl or halo.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from oneto 3. In certain instances, R¹³ is —CH₂OH. In certain instances, R¹³ isC₁₋₆ alkyl that is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is C₁₋₆ acyl. In certain instances, R¹³ is C₁acyl. In certain instances, R¹³ is C₂ acyl. In certain instances, R¹³ isC₃ acyl. In certain instances, R¹³ is C₄₋₆ acyl.

In certain instances, R¹³ is SO₂—C₁₋₆alkyl. In certain instances, R¹³ isSO₂—C₁alkyl. n certain instances, R¹³ is SO₂—C₂alkyl. In certaininstances, R¹³ is SO₂—C₃alkyl. In certain instances, R¹³ isSO₂—C₄₋₆alkyl.

In certain instances, R¹³ is C₃₋₇ cycloalkyl. In certain instances, R¹³is unsubstituted C₃ cycloalkyl. In certain instances, R¹³ isunsubstituted C₄ cycloalkyl. In certain instances, R¹³ is unsubstitutedC₅₋₆ cycloalkyl. In certain instances, R¹³ is unsubstituted C₇cycloalkyl.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is halo. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is fluoro.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹⁰ is methyl.In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is halo; and R¹⁰ ismethyl. In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is fluoro; andR¹⁰ is methyl.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ ishydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo;and R¹³ is hydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen;R⁹ is fluoro; and R¹³ is hydrogen.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is —CH₂OH.In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³is —CH₂OH. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ isfluoro; and R¹³ is —CH₂OH.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is—(CH₂)_(m)F, wherein m is a number from one to 3. In certain instances,R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³ is —(CH₂)_(m)F, whereinm is a number from one to 3. In certain instances, R⁶, R⁷, and R⁸ arehydrogen; R⁹ is fluoro; and R¹³ is —(CH₂)_(m)F, wherein m is a numberfrom one to 3.

Formula (II)

The present disclosure provides a compound of Formula (II):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²¹ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In Formula (II), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are hydrogen.

In Formula (II), n is a number from zero to 4. In certain instances, nis zero. In certain instances, n is one. In certain instances, n is 2.In certain instances, n is 3. In certain instances, n is 4.

In Formula (II), R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆alkoxy, cyano, and nitro. In certain instances, R³ is halo. In certaininstances, R³ is hydroxyl. In certain instances, R³ is C₁₋₆ alkyl. Incertain instances, R³ is C₁₋₆ alkoxy. In certain instances, R³ is cyano.In certain instances, R³ is nitro.

In Formula (II), R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇cycloalkyl, and —NR²²R²³; wherein the alkyl or cycloalkyl isunsubstituted or substituted with hydroxyl or amino; and wherein eachR²² and R²³ are independently selected from hydrogen and C₁₋₆ alkyl orR²² and R²¹ may be joined to form a 3 to 10 membered ring. In certaininstances, R⁴ is hydrogen. In certain instances, R⁴ is C₁₋₆ alkyl. Incertain instances, R⁴ is C₃₋₇ cycloalkyl. In certain instances, R⁴ is—NR²²R²³.

In certain instances, R⁴ is unsubstituted C₁₋₆ alkyl. In certaininstances, R⁴ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with hydroxyl.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with amino.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₆ alkyl that is substituted with —N(CH₃)₂.In certain instances, R⁴ is C₁₋₃ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with —N(CH₃)₂.

In certain instances, R⁴ is unsubstituted C₃₋₇ cycloalkyl. In certaininstances, R⁴ is unsubstituted C₃ cycloalkyl. In certain instances, R⁴is unsubstituted C₄ cycloalkyl. In certain instances, R⁴ isunsubstituted C₅₋₆ cycloalkyl. In certain instances, R⁴ is unsubstitutedC₇ cycloalkyl.

In certain instances, R⁴ is —NR²²R²³, wherein each R²² and R²³ areindependently selected from hydrogen and C₁₋₆ alkyl or R²² and R²³ maybe joined to form a 3 to 10 membered ring. In certain instances, R²² andR²³ are hydrogen. In certain instances, R²² and R²³ are C₁₋₆ alkyl. Incertain instances, R²² and R²³ are C₁₋₃ alkyl. In certain instances, R²²and R²³ are methyl.

In certain instances, R²² and R²³ may be joined to form a 3 to 10membered ring, such that R⁴ is

where w is a number from 1 to 8. In certain instances, R²² and R²³ maybe joined to form a 3-membered ring. In certain instances, R²² and R²³may be joined to form a 4-membered ring. In certain instances, R²² andR²³ may be joined to form a 5-membered ring.

In certain instances, R²² and R²³ may be joined to form a 6-memberedring. In certain instances, R²² and R²³ may be joined to form a7-membered ring.

In Formula (II), R⁵ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R⁵ is hydrogen. In certain instances, R⁵ is C₁₋₆ alkyl. Incertain instances, R⁵ is methyl. In certain instances, R⁵ is ethyl. Incertain instances, R⁵ is C₁₋₃ alkyl. In certain instances, R⁵ is C₄₋₆alkyl.

In Formula (II), R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁶ is hydrogen. In certain instances, R⁶ is halo. Incertain instances, R⁶ is fluoro. In certain instances, R⁶ is chloro. Incertain instances, R⁶ is bromo. In certain instances, R⁶ is C₁₋₆ alkyl.In certain instances, R⁶ is C₁₋₆ haloalkyl. In certain instances, R⁶ isC₂₋₆ alkoxy. In certain instances, R⁶ is C₁₋₆ haloalkoxy. In certaininstances, R⁶ is hydroxyl. In certain instances, R⁶ is cyano. In certaininstances, R⁶ is nitro.

In Formula (II), R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁷ is hydrogen. In certain instances, R⁷ is halo. Incertain instances, R⁷ is fluoro. In certain instances, R⁷ is chloro. Incertain instances, R⁷ is bromo. In certain instances, R⁷ is C₁₋₆ alkyl.In certain instances, R⁷ is C₁₋₆ haloalkyl. In certain instances, R⁷ isC₂₋₆ alkoxy. In certain instances, R⁷ is C₁₋₆ haloalkoxy. In certaininstances, R⁷ is hydroxyl. In certain instances, R⁷ is cyano. In certaininstances, R⁷ is nitro.

In Formula (II), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (II), Q is CR⁹ or N. In certain instances, Q is CR⁹. Incertain instances, Q is N.

In Formula (II), R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁹ is hydrogen. In certain instances, R⁹ is halo. Incertain instances, R⁹ is fluoro. In certain instances, R⁹ is chloro. Incertain instances, R⁹ is bromo. In certain instances, R⁹ is C₁₋₆ alkyl.In certain instances, R⁹ is C₁₋₆ haloalkyl. In certain instances, R⁹ isC₁₋₆ alkoxy. In certain instances, R⁹ is C₁₋₆ haloalkoxy. In certaininstances, R⁹ is hydroxyl. In certain instances, R⁹ is cyano. In certaininstances, R⁹ is nitro.

In Formula (II), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆alkyl. In certain instances, R¹⁰ is methyl. In certain instances, R¹⁰ isethyl. In certain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances,R¹⁰ is C₄₋₆ alkyl.

In Formula (II), R¹¹ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹¹ is hydrogen. In certain instances, R¹¹ is C₁₋₆alkyl. In certain instances, R¹¹ is methyl. In certain instances, R¹¹ isethyl. In certain instances, R¹¹ is C₁₋₃ alkyl. In certain instances,R¹¹ is C₄₋₆ alkyl.

In Formula (II), R¹² is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹² is hydrogen. In certain instances, R¹² is C₁₋₆alkyl. In certain instances, R¹² is methyl. In certain instances, R¹² isethyl. In certain instances, R¹² is C₁₋₃ alkyl. In certain instances,R¹² is C₄₋₆ alkyl.

In Formula (II), R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀aryl, wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo. In certain instances, R¹³ is hydrogen.In certain instances, R¹³ is C₁₋₆ alkyl. In certain instances, R¹³ isC₆₋₂₀ aryl.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from oneto 3. In certain instances, R¹³ is —CH₂OH. In certain instances, R¹³ isC₁₋₆ alkyl that is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is halo. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is fluoro.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹⁰ is methyl.In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is halo; and R¹⁰ ismethyl. In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is fluoro; andR¹⁰ is methyl.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ ishydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo;and R¹³ is hydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen;R⁹ is fluoro; and R¹³ is hydrogen.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is —CH₂OH.In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³is —CH₂OH. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ isfluoro; and R¹³ is —CH₂OH.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is—(CH₂)_(m)F, wherein m is a number from one to 3. In certain instances,R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³ is —(CH₂)_(m)F, whereinm is a number from one to 3. In certain instances, R⁶, R⁷, and R⁸ arehydrogen; R⁹ is fluoro; and R¹³ is —(CH₂)_(m)F, wherein m is a numberfrom one to 3.

Formula (III)

The present disclosure provides a compound of Formula (III):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and R¹³ is selected fromhydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein each alkyl or aryl isunsubstituted or substituted with hydroxyl, C₁₋₆ alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In Formula (III), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are hydrogen.

In Formula (III), n is a number from zero to 4. In certain instances, nis zero. In certain instances, n is one. In certain instances, n is 2.In certain instances, n is 3. In certain instances, n is 4.

In Formula (III), R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆alkoxy, cyano, and nitro. In certain instances, R³ is halo. In certaininstances, R³ is hydroxyl. In certain instances, R³ is C₁₋₆ alkyl. Incertain instances, R³ is C₁₋₆ alkoxy. In certain instances, R³ is cyano.In certain instances, R³ is nitro.

In Formula (III), R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇cycloalkyl, and —NR²²R²³; wherein the alkyl or cycloalkyl isunsubstituted or substituted with hydroxyl or amino; and wherein eachR²² and R²³ are independently selected from hydrogen and C₁₋₆ alkyl orR²² and R²¹ may be joined to form a 3 to 10 membered ring. In certaininstances, R⁴ is hydrogen. In certain instances, R⁴ is C₁₋₆ alkyl. Incertain instances, R⁴ is C₃₋₇ cycloalkyl. In certain instances, R⁴ is—NR²²R²³.

In certain instances, R⁴ is unsubstituted C₁₋₆ alkyl. In certaininstances, R⁴ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with hydroxyl.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with amino.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₆ alkyl that is substituted with —N(CH₃)₂.In certain instances, R⁴ is C₁₋₃ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with —N(CH₃)₂.

In certain instances, R⁴ is unsubstituted C₃₋₇ cycloalkyl. In certaininstances, R⁴ is unsubstituted C₃ cycloalkyl. In certain instances, R⁴is unsubstituted C₄ cycloalkyl. In certain instances, R⁴ isunsubstituted C₅₋₆ cycloalkyl. In certain instances, R⁴ is unsubstitutedC₇ cycloalkyl.

In certain instances, R⁴ is —NR²²R²³, wherein each R²² and R²³ areindependently selected from hydrogen and C₁₋₆ alkyl or R²² and R²³ maybe joined to form a 3 to 10 membered ring. In certain instances, R²² andR²³ are hydrogen. In certain instances, R²² and R²³ are C₁₋₆ alkyl. Incertain instances, R²² and R²³ are C₁₋₃ alkyl. In certain instances, R²²and R²³ are methyl.

In certain instances, R²² and R²³ may be joined to form a 3 to 10membered ring, such that R⁴ is

where w is a number from 1 to 8. In certain instances, R²² and R²³ maybe joined to form a 3-membered ring. In certain instances, R²² and R²³may be joined to form a 4-membered ring. In certain instances, R²² andR²³ may be joined to form a 5-membered ring. In certain instances, R²²and R²³ may be joined to form a 6-membered ring. In certain instances,R²² and R²³ may be joined to form a 7-membered ring.

In Formula (III), R⁵ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R⁵ is hydrogen. In certain instances, R⁵ is C₁₋₆alkyl. In certain instances, R⁵ is methyl. In certain instances, R⁵ isethyl. In certain instances, R⁵ is C₁₋₃ alkyl. In certain instances, R⁵is C₄₋₆ alkyl.

In Formula (III), R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁶ is hydrogen. In certain instances, R⁶ is halo. Incertain instances, R⁶ is fluoro. In certain instances, R⁶ is chloro. Incertain instances, R⁶ is bromo. In certain instances, R⁶ is C₁₋₆ alkyl.In certain instances, R⁶ is C₁₋₆ haloalkyl. In certain instances, R⁶ isC₂₋₆ alkoxy. In certain instances, R⁶ is C₁₋₆ haloalkoxy. In certaininstances, R⁶ is hydroxyl. In certain instances, R⁶ is cyano. In certaininstances, R⁶ is nitro.

In Formula (III), R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁷ is hydrogen. In certain instances, R⁷ is halo. Incertain instances, R⁷ is fluoro. In certain instances, R⁷ is chloro. Incertain instances, R⁷ is bromo. In certain instances, R⁷ is C₁₋₆ alkyl.In certain instances, R⁷ is C₁₋₆ haloalkyl. In certain instances, R⁷ isC₂₋₆ alkoxy. In certain instances, R⁷ is C₁₋₆ haloalkoxy. In certaininstances, R⁷ is hydroxyl. In certain instances, R⁷ is cyano. In certaininstances, R⁷ is nitro.

In Formula (III), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (III), R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁹ is hydrogen. In certain instances, R⁹ is halo. Incertain instances, R⁹ is fluoro. In certain instances, R⁹ is chloro. Incertain instances, R⁹ is bromo. In certain instances, R⁹ is C₁₋₆ alkyl.In certain instances, R⁹ is C₁₋₆ haloalkyl. In certain instances, R⁹ isC₁₋₆ alkoxy. In certain instances, R⁹ is C₁₋₆ haloalkoxy. In certaininstances, R⁹ is hydroxyl. In certain instances, R⁹ is cyano. In certaininstances, R⁹ is nitro.

In Formula (III), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆alkyl. In certain instances, R¹⁰ is methyl. In certain instances, R¹⁰ isethyl. In certain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances,R¹⁰ is C₄₋₆ alkyl.

In Formula (III), R¹¹ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹¹ is hydrogen. In certain instances, R¹¹ is C₁₋₆alkyl. In certain instances, R¹¹ is methyl. In certain instances, R¹¹ isethyl. In certain instances, R¹¹ is C₁₋₃ alkyl. In certain instances,R¹¹ is C₄₋₆ alkyl.

In Formula (III), R¹² is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹² is hydrogen. In certain instances, R¹² is C₁₋₆alkyl. In certain instances, R¹² is methyl. In certain instances, R¹² isethyl. In certain instances, R¹² is C₁₋₃ alkyl. In certain instances,R¹² is C₄₋₆ alkyl.

In Formula (III), R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀aryl, wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo. In certain instances, R¹³ is hydrogen.In certain instances, R¹³ is C₁₋₆ alkyl. In certain instances, R¹³ isC₆₋₂₀ aryl.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is 6 alkyl that is substituted with hydroxyl. In certaininstances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from one to 3. Incertain instances, R¹³ is —CH₂OH. In certain instances, R¹³ is alkylthat is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is halo. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is fluoro.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹⁰ is methyl.In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is halo; and R¹⁰ ismethyl. In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is fluoro; andR¹⁰ is methyl.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ ishydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo;and R¹³ is hydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen;R⁹ is fluoro; and R¹³ is hydrogen.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is —CH₂OH.In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³is —CH₂OH. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ isfluoro; and R¹³ is —CH₂OH.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is—(CH₂)_(m)F, wherein m is a number from one to 3. In certain instances,R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³ is —(CH₂)_(m)F, whereinm is a number from one to 3. In certain instances, R⁶, R⁷, and R⁸ arehydrogen; R⁹ is fluoro; and R¹³ is —(CH₂)_(m)F, wherein m is a numberfrom one to 3.

Formula (IV)

The present disclosure provides a compound of Formula (IV):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy, cyano, andnitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;

-   -   wherein the alkyl or cycloalkyl is unsubstituted or substituted        with hydroxyl or amino; and    -   wherein each R²² and R²³ are independently selected from        hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined to form a 3        to 10 membered ring;

R⁵ is selected from hydrogen and C₁₋₆ alkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In Formula (IV), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are hydrogen.

In Formula (IV), n is a number from zero to 4. In certain instances, nis zero. In certain instances, n is one. In certain instances, n is 2.In certain instances, n is 3. In certain instances, n is 4.

In Formula (IV), R³ is selected from halo, hydroxyl, C₁₋₆ alkyl, C₁₋₆alkoxy, cyano, and nitro. In certain instances, R³ is halo. In certaininstances, R³ is hydroxyl. In certain instances, R³ is C₁₋₆ alkyl. Incertain instances, R³ is C₁₋₆ alkoxy. In certain instances, R³ is cyano.In certain instances, R³ is nitro.

In Formula (IV), R⁴ is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇cycloalkyl, and —NR²²R²³; wherein the alkyl or cycloalkyl isunsubstituted or substituted with hydroxyl or amino; and wherein eachR²² and R²³ are independently selected from hydrogen and C₁₋₆ alkyl orR²² and R²¹ may be joined to form a 3 to 10 membered ring. In certaininstances, R⁴ is hydrogen. In certain instances, R⁴ is C₁₋₆ alkyl. Incertain instances, R⁴ is C₃₋₇ cycloalkyl. In certain instances, R⁴ is—NR²²R²³.

In certain instances, R⁴ is unsubstituted C₁₋₆ alkyl. In certaininstances, R⁴ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with hydroxyl.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with amino.In certain instances, R⁴ is C₁₋₆ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₆ alkyl that is substituted with —N(CH₃)₂.In certain instances, R⁴ is C₁₋₃ alkyl that is substituted with —NH₂. Incertain instances, R⁴ is C₁₋₃ alkyl that is substituted with —N(CH₃)₂.

In certain instances, R⁴ is unsubstituted C₃₋₇ cycloalkyl. In certaininstances, R⁴ is unsubstituted C₃ cycloalkyl. In certain instances, R⁴is unsubstituted C₄ cycloalkyl. In certain instances, R⁴ isunsubstituted C₅₋₆ cycloalkyl. In certain instances, R⁴ is unsubstitutedC₇ cycloalkyl.

In certain instances, R⁴ is —NR²²R²³, wherein each R²² and R²³ areindependently selected from hydrogen and C₁₋₆ alkyl or R²² and R²³ maybe joined to form a 3 to 10 membered ring. In certain instances, R²² andR²³ are hydrogen. In certain instances, R²² and R²³ are C₁₋₆ alkyl. Incertain instances, R²² and R²³ are C₁₋₃ alkyl. In certain instances, R²²and R²³ are methyl.

In certain instances, R²² and R²³ may be joined to form a 3 to 10membered ring, such that R⁴ is

where w is a number from 1 to 8. In certain instances, R²² and R²³ maybe joined to form a 3-membered ring. In certain instances, R²² and R²³may be joined to form a 4-membered ring. In certain instances, R²² andR²³ may be joined to form a 5-membered ring. In certain instances, R²²and R²³ may be joined to form a 6-membered ring. In certain instances,R²² and R²³ may be joined to form a 7-membered ring.

In Formula (IV), R⁵ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R⁵ is hydrogen. In certain instances, R⁵ is C₁₋₆ alkyl. Incertain instances, R⁵ is methyl. In certain instances, R⁵ is ethyl. Incertain instances, R⁵ is C₁₋₃ alkyl. In certain instances, R⁵ is C₄₋₆alkyl.

In Formula (IV), R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁶ is hydrogen. In certain instances, R⁶ is halo. Incertain instances, R⁶ is fluoro. In certain instances, R⁶ is chloro. Incertain instances, R⁶ is bromo. In certain instances, R⁶ is C₁₋₆ alkyl.In certain instances, R⁶ is C₁₋₆ haloalkyl. In certain instances, R⁶ isC₂₋₆ alkoxy. In certain instances, R⁶ is C₁₋₆ haloalkoxy. In certaininstances, R⁶ is hydroxyl. In certain instances, R⁶ is cyano. In certaininstances, R⁶ is nitro.

In Formula (IV), R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁷ is hydrogen. In certain instances, R⁷ is halo. Incertain instances, R⁷ is fluoro. In certain instances, R⁷ is chloro. Incertain instances, R⁷ is bromo. In certain instances, R⁷ is C₁₋₆ alkyl.In certain instances, R⁷ is C₁₋₆ haloalkyl. In certain instances, R⁷ isC₂₋₆ alkoxy. In certain instances, R⁷ is C₁₋₆ haloalkoxy. In certaininstances, R⁷ is hydroxyl. In certain instances, R⁷ is cyano. In certaininstances, R⁷ is nitro.

In Formula (IV), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (IV), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆alkyl. In certain instances, R¹⁰ is methyl. In certain instances, R¹⁰ isethyl. In certain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances,R¹⁰ is C₄₋₆ alkyl.

In Formula (IV), R¹¹ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹² is hydrogen. In certain instances, R¹¹ is C₁₋₆alkyl. In certain instances, R¹¹ is methyl. In certain instances, R¹¹ isethyl. In certain instances, R¹¹ is C₁₋₃ alkyl. In certain instances,R¹¹ is C₄₋₆ alkyl.

In Formula (IV), R¹² is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹² is hydrogen. In certain instances, R¹² is C₁₋₆alkyl. In certain instances, R¹² is methyl. In certain instances, R¹² isethyl. In certain instances, R¹² is C₁₋₃ alkyl. In certain instances,R¹² is C₄₋₆ alkyl.

In Formula (IV), R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀aryl, wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo. In certain instances, R¹³ is hydrogen.In certain instances, R¹³ is C₁₋₆ alkyl. In certain instances, R¹³ isC₆₋₂₀ aryl.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from oneto 3. In certain instances, R¹³ is —CH₂OH. In certain instances, R¹³ isC₁₋₆ alkyl that is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁶, R⁷, and R⁸ are hydrogen. In certain instances,R⁶, R⁷, and R⁸ are hydrogen and R¹⁰ is methyl. In certain instances, R⁶,R⁷, and R⁸ are hydrogen and R¹³ is hydrogen. In certain instances, R⁶,R⁷, and R⁸ are hydrogen and R¹³ is —CH₂OH. In certain instances, R⁶, R⁷,and R⁸ are hydrogen and R¹³ is hydrogen. In certain instances, R⁶, R⁷,and R⁸ are hydrogen and R¹³ is —(CH₂)_(m)F, wherein m is a number fromone to 3.

Formula (V)

The present disclosure provides a compound of Formula (V):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl;

R¹¹ is selected from hydrogen and C₁₋₆ alkyl;

R¹² is selected from hydrogen and C₁₋₆ alkyl;

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In Formula (V), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are hydrogen.

In Formula (V), R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁶ is hydrogen. In certain instances, R⁶ is halo. Incertain instances, R⁶ is fluoro. In certain instances, R⁶ is chloro. Incertain instances, R⁶ is bromo. In certain instances, R⁶ is C₁₋₆ alkyl.In certain instances, R⁶ is C₁₋₆ haloalkyl. In certain instances, R⁶ isC₂₋₆ alkoxy. In certain instances, R⁶ is C₁₋₆ haloalkoxy. In certaininstances, R⁶ is hydroxyl. In certain instances, R⁶ is cyano. In certaininstances, R⁶ is nitro.

In Formula (V), R⁷ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁷ is hydrogen. In certain instances, R⁷ is halo. Incertain instances, R⁷ is fluoro. In certain instances, R⁷ is chloro. Incertain instances, R⁷ is bromo. In certain instances, R⁷ is C₁₋₆ alkyl.In certain instances, R⁷ is C₁₋₆ haloalkyl. In certain instances, R⁷ isC₂₋₆ alkoxy. In certain instances, R⁷ is C₁₋₆ haloalkoxy. In certaininstances, R⁷ is hydroxyl. In certain instances, R⁷ is cyano. In certaininstances, R⁷ is nitro.

In Formula (V), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (V), Q is CR⁹ or N. In certain instances, Q is CR⁹. Incertain instances, Q is N.

In Formula (V), R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁹ is hydrogen. In certain instances, R⁹ is halo. Incertain instances, R⁹ is fluoro. In certain instances, R⁹ is chloro. Incertain instances, R⁹ is bromo. In certain instances, R⁹ is C₁₋₆ alkyl.In certain instances, R⁹ is C₁₋₆ haloalkyl. In certain instances, R⁹ isC₁₋₆ alkoxy. In certain instances, R⁹ is C₁₋₆ haloalkoxy. In certaininstances, R⁹ is hydroxyl. In certain instances, R⁹ is cyano. In certaininstances, R⁹ is nitro.

In Formula (V), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆ alkyl. Incertain instances, R¹⁰ is methyl. In certain instances, R¹⁰ is ethyl. Incertain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances, R¹⁰ is C₄₋₆alkyl.

In Formula (V), R¹¹ is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹¹ is hydrogen. In certain instances, R¹¹ is C₁₋₆ alkyl. Incertain instances, R¹¹ is methyl. In certain instances, R¹¹ is ethyl. Incertain instances, R¹¹ is C₁₋₃ alkyl. In certain instances, R¹¹ is C₄₋₆alkyl.

In Formula (V), R¹² is selected from hydrogen and C₁₋₆ alkyl. In certaininstances, R¹² is hydrogen. In certain instances, R¹² is C₁₋₆ alkyl. Incertain instances, R¹² is methyl. In certain instances, R¹² is ethyl. Incertain instances, R¹² is C₁₋₃ alkyl. In certain instances, R¹² is C₄₋₆alkyl.

In Formula (V), R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀aryl, wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo. In certain instances, R¹³ is hydrogen.In certain instances, R¹³ is C₁₋₆ alkyl. In certain instances, R¹³ isC₆₋₂₀ aryl.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from oneto 3. In certain instances, R¹³ is —CH₂OH. In certain instances, R¹³ isC₁₋₆ alkyl that is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is halo. In certaininstances, R⁶, R⁷, and R⁸ are hydrogen and R⁹ is fluoro.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹⁰ is methyl.In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is halo; and R¹⁰ ismethyl. In certain instances, R⁶, R⁷, R⁸ are hydrogen; R⁹ is fluoro; andR¹⁰ is methyl.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ ishydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo;and R¹³ is hydrogen. In certain instances, R⁶, R⁷, and R⁸ are hydrogen;R⁹ is fluoro; and R¹³ is hydrogen.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is —CH₂OH.In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³is —CH₂OH. In certain instances, R⁶, R⁷, and R⁸ are hydrogen; R⁹ isfluoro; and R¹³ is —CH₂OH.

In certain instances, R⁶, R⁷, R⁸, and R⁹ are hydrogen and R¹³ is—(CH₂)_(m)F, wherein m is a number from one to 3. In certain instances,R⁶, R⁷, and R⁸ are hydrogen; R⁹ is halo; and R¹³ is —(CH₂)_(m)F, whereinm is a number from one to 3. In certain instances, R⁶, R⁷, and R⁸ arehydrogen; R⁹ is fluoro; and R¹³ is —(CH₂)_(m)F, wherein m is a numberfrom one to 3.

Formula (VI)

The present disclosure provides a compound of Formula (VI):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is selected from hydrogen and C₁₋₆ alkyl; and

R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein eachalkyl or aryl is unsubstituted or substituted with hydroxyl, C₁₋₆alkoxy, or halo;

or a pharmaceutically acceptable salt thereof.

In Formula (VI), R¹ and R² are independently selected from hydrogen,halo, C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In certain instances, R¹ ishydrogen. In certain instances, R¹ is C₁₋₆ alkyl. In certain instances,R¹ is methyl or ethyl. In certain instances, R² is hydrogen. In certaininstances, R² is C₁₋₆ alkyl. In certain instances, R² is methyl orethyl. In certain instances, R¹ and R² are hydrogen.

In Formula (VI), R⁸ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁸ is hydrogen. In certain instances, R⁸ is halo. Incertain instances, R⁸ is fluoro. In certain instances, R⁸ is chloro. Incertain instances, R⁸ is bromo. In certain instances, R⁸ is C₁₋₆ alkyl.In certain instances, R⁸ is C₁₋₆ haloalkyl. In certain instances, R⁸ isC₁₋₆ alkoxy. In certain instances, R⁸ is C₁₋₆ haloalkoxy. In certaininstances, R⁸ is hydroxyl. In certain instances, R⁸ is cyano. In certaininstances, R⁸ is nitro.

In Formula (VI), Q is CR⁹ or N. In certain instances, Q is CR⁹. Incertain instances, Q is N.

In Formula (VI), R⁹ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro. Incertain instances, R⁹ is hydrogen. In certain instances, R⁹ is halo. Incertain instances, R⁹ is fluoro. In certain instances, R⁹ is chloro. Incertain instances, R⁹ is bromo. In certain instances, R⁹ is C₁₋₆ alkyl.In certain instances, R⁹ is C₁₋₆ haloalkyl. In certain instances, R⁹ isC₁₋₆ alkoxy. In certain instances, R⁹ is C₁₋₆ haloalkoxy. In certaininstances, R⁹ is hydroxyl. In certain instances, R⁹ is cyano. In certaininstances, R⁹ is nitro.

In Formula (VI), R¹⁰ is selected from hydrogen and C₁₋₆ alkyl. Incertain instances, R¹⁰ is hydrogen. In certain instances, R¹⁰ is C₁₋₆alkyl. In certain instances, R¹⁰ is methyl. In certain instances, R¹⁰ isethyl. In certain instances, R¹⁰ is C₁₋₃ alkyl. In certain instances,R¹⁰ is C₄₋₆ alkyl.

In Formula (VI), R¹³ is selected from hydrogen, C₁₋₆ alkyl, and C₆₋₂₀aryl, wherein each alkyl or aryl is unsubstituted or substituted withhydroxyl, C₁₋₆ alkoxy, or halo. In certain instances, R¹³ is hydrogen.In certain instances, R¹³ is C₁₋₆ alkyl. In certain instances, R¹³ isC₆₋₂₀ aryl.

In certain instances, R¹³ is unsubstituted C₁₋₆ alkyl. In certaininstances, R¹³ is C₁₋₆ alkyl that is substituted with hydroxyl. Incertain instances, R¹³ is —(CH₂)_(m)OH, wherein m is a number from oneto 3. In certain instances, R¹³ is —CH₂OH. In certain instances, R¹³ isC₁₋₆ alkyl that is substituted with C₁₋₆ alkoxy.

In certain instances, R¹³ is C₁₋₆ alkyl that is substituted with halo.In certain instances, R¹³ is —(CH₂)_(m)X, wherein m is a number from oneto 3 and X is halo. In certain instances, R¹³ is C₁₋₆ alkyl that issubstituted with fluoro. In certain instances, R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R¹³ is—(CH₂)₂F.

In certain instances, R¹³ is unsubstituted C₆₋₂₀ aryl. In certaininstances, R¹³ is C₆₋₂₀ aryl that is substituted with hydroxyl. Incertain instances, R¹³ is C₆₋₂₀ aryl that is substituted with C₁₋₆alkoxy. In certain instances, R¹³ is C₆₋₂₀ aryl that is substituted withhalo.

In certain instances, R⁸ and R⁹ are hydrogen. In certain instances, R⁸is hydrogen and R⁹ is halo. In certain instances, R⁸ is hydrogen and R⁹is fluoro.

In certain instances, R⁸ and R⁹ are hydrogen and R¹⁰ is methyl. Incertain instances, R⁸ is hydrogen; R⁹ is halo; and R¹⁰ is methyl. Incertain instances, R⁸ is hydrogen; R⁹ is fluoro; and R¹⁰ is methyl.

In certain instances, R⁸ and R⁹ are hydrogen and R¹³ is hydrogen. Incertain instances, R⁸ is hydrogen; R⁹ is halo; and R¹³ is hydrogen. Incertain instances, R⁸ is hydrogen; R⁹ is fluoro; and R¹³ is hydrogen.

In certain instances, R⁸ and R⁹ are hydrogen and R¹³ is —CH₂OH. Incertain instances, R⁸ is hydrogen; R⁹ is halo; and R¹³ is —CH₂OH. Incertain instances, R⁸ is hydrogen; R⁹ is fluoro; and R¹³ is —CH₂OH.

In certain instances, R⁸ and R⁹ are hydrogen and R¹³ is —(CH₂)_(m)F,wherein m is a number from one to 3. In certain instances, R⁸ ishydrogen; R⁹ is halo; and R¹³ is —(CH₂)_(m)F, wherein m is a number fromone to 3. In certain instances, R⁸ is hydrogen; R⁹ is fluoro; and R¹³ is—(CH₂)_(m)F, wherein m is a number from one to 3.

Formula (VII)

The present disclosure provides a compound of Formula (VII):

wherein

R¹ and R² are independently selected from hydrogen, halo, C₁₋₆ alkyl,and C₁₋₆ haloalkyl;

R⁸ is selected from halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, hydroxyl, cyano, and nitro;

R¹⁰ is C₁₋₆ alkyl; and

R¹³ is hydrogen or C₁₋₆ alkyl;

or a pharmaceutically acceptable salt thereof.

In some embodiments of Formula (VII), R¹ and R² are each hydrogen. Inother embodiments, R⁸ is halo, methyl, methoxy, or cyano. In still otherembodiments, R⁸ is halo. In still other embodiments, R⁸ is fluoro. Insome embodiments, R¹⁰ is methyl, ethyl, or isopropyl. In otherembodiments, R¹⁰ is methyl. In some embodiments, R¹³ is hydrogen,methyl, ethyl, or isopropyl. In other embodiments, R¹³ is hydrogen.

In some embodiments of compounds of Formulae (I)-(VI), R⁶ and R⁷ mayalso be methoxy, provided that neither R⁶ nor R⁷ is methoxy when R¹⁰ ismethyl.

In certain embodiments, the present disclosure provides a compoundselected from:

Compound Structure Chemical Name 1

N-(3-((2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin- 4-yl)oxy)phenyl)acrylamide2

N-(3-((2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4- yl)oxy)phenyl)acrylamide 3

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin- 4-yl)oxy)phenyl)acrylamide4

N-(3-((7-(hydroxymethyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 5

N-(3-((7-(hydroxymethyl)-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 6

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7-(hydroxymethyl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide; and 7

N-(3-((7-(2-fluoroethyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamideand pharmaceutically acceptable salts thereof.

In certain embodiments, the present disclosure provides a compoundselected from:

Compound Structure Chemical Name 8

N-(3-((2-((1-(2-(dimethylamino)ethyl)-1H-indol-5-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin- 4-yl)oxy)phenyl)acrylamide9

N-(3-((2-((2-((dimethylamino)methyl)quinolin-6-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4- yl)oxy)phenyl)acrylamide 10

N-(3-((5-cyclopropyl-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 11

N-(3((5-cyclopropyl-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 12

N-(3-((2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(pyrrolidin-1-yl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 13

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(pyrrolidin-1-yl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 14

N-(3-((5-(2-hydroxyethyl)-2((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 15

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(2-hydroxyethyl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 16

N-(3-((5-((dimethylamino)methyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 17

N-(3-((5-((dimethylamino)methyl)-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 18

N-(3((5-(dimethylamino)-2((4-(4- methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4- yl)oxy)phenyl)acrylamide 19

N-(3-((5-(dimethylamino)-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 20

N-(3-((5-(2-(dimethylamino)ethyl)-2((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 21

N-(3-((5-(2-(dimethylamino)ethyl)-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 22

N-(3-((5-(aziridin-1-yl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 23

N-(3-((5-(aziridin-1-yl)-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 24

N-(3-((5-(azetidin-1-yl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 25

N-(3-((5-(azetidin-1-yl)-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 26

N-(3-((2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(piperidin-1-yl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 27

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(piperidin-1-yl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 28

N-(3-((7-cyclopropyl-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 29

N-(3-((7-cyclopropyl-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 30

N-(3-((2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7-(methylsulfonyl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 31

N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7-(methylsulfonyl)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 32

N-(3-((7-acetyl-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin- 4-yl)oxy)phenyl)acrylamide33

N-(3-((7-acetyl-2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H- pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide 34

N-(3-(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-7H- pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide 35

N-(3-(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-7H- pyrrolo[2,3-d]pyrimidin-4-ylamino)phenyl)acrylamide; and 36

N-(3-((2-((2-(piperidin-1-ylmethyl)quinolin-6-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4- yl)oxy)phenyl)acrylamideand pharmaceutically acceptable salts thereof.

In certain embodiments, the present disclosure provides Compound 3,N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide,

and pharmaceutically acceptable salts thereof. In certain embodiments,the present disclosure provides the maleate salt of Compound 3,N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide.In certain embodiments, the present disclosure provides thehydrochloride salt of Compound 3,N-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide.

The disclosed pharmaceutical compositions can be formulated as apharmaceutically acceptable salt of a disclosed compound.Pharmaceutically acceptable salts are non-toxic salts of a free baseform of a compound that possesses the desired pharmacological activityof the free base. These salts may be derived from inorganic or organicacids. Non-limiting examples of pharmaceutically acceptable saltsinclude sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,phosphates, monohydrogen-phosphates, dihydrogenphosphates,metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates,propionates, decanoates, caprylates, acrylates, formates, isobutyrates,caproates, heptanoates, propiolates, oxalates, malonates, succinates,suberates, sebacates, fumarates, maleates, butyne-1,4-dioates,hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates,sulfonates, methylsulfonates, propylsulfonates, besylates,xylenesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates,phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates,γ-hydroxybutyrates, glycolates, tartrates, and mandelates. Lists ofother suitable pharmaceutically acceptable salts are found inRemington's Pharmaceutical Sciences, 17th Edition, Mack PublishingCompany, Easton, Pa., 1985.

Pharmaceutical Compositions

For treatment purposes, pharmaceutical compositions comprising thecompounds described herein may further comprise one or morepharmaceutically-acceptable excipients. A pharmaceutically-acceptableexcipient is a substance that is non-toxic and otherwise biologicallysuitable for administration to a subject. Such excipients facilitateadministration of the compounds described herein and are compatible withthe active ingredient. Examples of pharmaceutically-acceptableexcipients include stabilizers, lubricants, surfactants, diluents,anti-oxidants, binders, coloring agents, bulking agents, emulsifiers, ortaste-modifying agents. In preferred embodiments, pharmaceuticalcompositions according to the embodiments are sterile compositions.Pharmaceutical compositions may be prepared using compounding techniquesknown or that become available to those skilled in the art.

Sterile compositions are within the present disclosure, includingcompositions that are in accord with national and local regulationsgoverning such compositions.

The pharmaceutical compositions and compounds described herein may beformulated as solutions, emulsions, suspensions, or dispersions insuitable pharmaceutical solvents or carriers, or as pills, tablets,lozenges, suppositories, sachets, dragees, granules, powders, powdersfor reconstitution, or capsules along with solid carriers according toconventional methods known in the art for preparation of various dosageforms. Pharmaceutical compositions of the embodiments may beadministered by a suitable route of delivery, such as oral, parenteral,rectal, nasal, topical, or ocular routes, or by inhalation. Preferably,the compositions are formulated for intravenous or oral administration.

For oral administration, the compounds of the embodiments may beprovided in a solid form, such as a tablet or capsule, or as a solution,emulsion, or suspension. To prepare the oral compositions, the compoundsof the embodiments may be formulated to yield a dosage of, e.g., fromabout 0.01 to about 50 mg/kg daily, or from about 0.05 to about 20 mg/kgdaily, or from about 0.1 to about 10 mg/kg daily. Oral tablets mayinclude the active ingredient(s) mixed with compatible pharmaceuticallyacceptable excipients such as diluents, disintegrating agents, bindingagents, lubricating agents, sweetening agents, flavoring agents,coloring agents and preservative agents. Suitable inert fillers includesodium and calcium carbonate, sodium and calcium phosphate, lactose,starch, sugar, glucose, methyl cellulose, magnesium stearate, mannitol,sorbitol, and the like. Exemplary liquid oral excipients includeethanol, glycerol, water, and the like. Starch, polyvinyl-pyrrolidone(PVP), sodium starch glycolate, microcrystalline cellulose, and alginicacid are exemplary disintegrating agents. Binding agents may includestarch and gelatin. The lubricating agent, if present, may be magnesiumstearate, stearic acid, or talc. If desired, the tablets may be coatedwith a material such as glyceryl monostearate or glyceryl distearate todelay absorption in the gastrointestinal tract, or may be coated with anenteric coating.

Capsules for oral administration include hard and soft gelatin capsules.To prepare hard gelatin capsules, active ingredient(s) may be mixed witha solid, semi-solid, or liquid diluent. Soft gelatin capsules may beprepared by mixing the active ingredient with water, an oil, such aspeanut oil or olive oil, liquid paraffin, a mixture of mono anddi-glycerides of short chain fatty acids, polyethylene glycol 400, orpropylene glycol.

Liquids for oral administration may be in the form of suspensions,solutions, emulsions, or syrups, or may be lyophilized or presented as adry product for reconstitution with water or other suitable vehiclebefore use. Such liquid compositions may optionally contain:pharmaceutically-acceptable excipients such as suspending agents (forexample, sorbitol, methyl cellulose, sodium alginate, gelatin,hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel andthe like); non-aqueous vehicles, e.g., oil (for example, almond oil orfractionated coconut oil), propylene glycol, ethyl alcohol, or water;preservatives (for example, methyl or propyl p-hydroxybenzoate or sorbicacid); wetting agents such as lecithin; and, if desired, flavoring orcoloring agents.

The compositions may be formulated for rectal administration as asuppository. For parenteral use, including intravenous, intramuscular,intraperitoneal, intranasal, or subcutaneous routes, the compounds ofthe embodiments may be provided in sterile aqueous solutions orsuspensions, buffered to an appropriate pH and isotonicity or inparenterally acceptable oil. Suitable aqueous vehicles include Ringer'ssolution and isotonic sodium chloride. Such forms may be presented inunit-dose form such as ampoules or disposable injection devices, inmulti-dose forms such as vials from which the appropriate dose may bewithdrawn, or in a solid form or pre-concentrate that can be used toprepare an injectable formulation. Illustrative infusion doses rangefrom about 1 to 1000 μg/kg/minute of agent admixed with a pharmaceuticalcarrier over a period ranging from several minutes to several days.

For nasal, inhaled, or oral administration, the pharmaceuticalcompositions may be administered using, for example, a spray formulationalso containing a suitable carrier.

For topical applications, the compounds of the embodiments arepreferably formulated as creams or ointments or a similar vehiclesuitable for topical administration. For topical administration, thecompounds of the embodiments may be mixed with a pharmaceutical carrierat a concentration of about 0.1% to about 10% of drug to vehicle.Another mode of administering the compounds of the embodiments mayutilize a patch formulation to effect transdermal delivery.

In certain embodiments, the present disclosure provides pharmaceuticalcomposition comprising a compound of formulae (I)-(VIII) andmethylcellulose. In certain embodiments, methylcellulose is in asuspension of about 0.1, 0.2, 0.3, 0.4, or 0.5 to about 1%. In certainembodiments, methylcellulose is in a suspension of about 0.1 to about0.5, 0.6, 0.7, 0.8, 0.9, or 1%. In certain embodiments, methylcelluloseis in a suspension of about 0.1 to about 1%. In certain embodiments,methylcellulose is in a suspension of about 0.1, 0.2, 0.3, 0.4, 0.5,0.6, 0.7, 0.8, 0.8, or 1%. In certain embodiments, methylcellulose is ina suspension of about 0.5%.

As used herein, the terms “treat” or “treatment” encompass both“preventative” and “curative” treatment. “Preventative” treatment ismeant to indicate a postponement of development of a disease, a symptomof a disease, or medical condition, suppressing symptoms that mayappear, or reducing the risk of developing or recurrence of a disease orsymptom. “Curative” treatment includes reducing the severity of orsuppressing the worsening of an existing disease, symptom, or condition.Thus, treatment includes ameliorating or preventing the worsening ofexisting disease symptoms, preventing additional symptoms fromoccurring, ameliorating or preventing the underlying systemic causes ofsymptoms, inhibiting the disorder or disease, e.g., arresting thedevelopment of the disorder or disease, relieving the disorder ordisease, causing regression of the disorder or disease, relieving acondition caused by the disease or disorder, or stopping the symptoms ofthe disease or disorder.

One of ordinary skill in the art may modify the formulations within theteachings of the specification to provide numerous formulations for aparticular route of administration. In particular, the compounds may bemodified to render them more soluble in water or other vehicle. It isalso well within the ordinary skill of the art to modify the route ofadministration and dosage regimen of a particular compound in order tomanage the pharmacokinetics of the present compounds for maximumbeneficial effect in a patient.

The term “subject” refers to a mammalian patient in need of suchtreatment, such as a human.

The compounds can be administered to a subject in need of treatment fora proliferation disorder. An example of a proliferation disorder iscancer. In certain instances, the compounds are useful to treat sarcoma,epidermoid cancer, fibrosarcoma, cervical cancer, gastric carcinoma,skin cancer, leukemia, lymphoma, lung cancer, non-small cell lungcancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renalcancer, prostate cancer, breast cancer, liver cancer, head and neckcancers, and pancreatic cancer.

In one aspect, the compounds and pharmaceutical compositions of theembodiments can be administered to a subject in need of treatment of acondition associated with EGFR inhibitory activity targetingsubstantially a mutated EGFR but not substantially the wild type EGFR.In some embodiments, the mutated EGFR comprises a T790M mutation. Thepresent disclosure provides use of a compound of Formulae (I)-(VIII) inthe preparation of a medicament for the treatment of such conditions,and the use of such compounds and salts for treatment of suchconditions.

In another aspect, the present disclosure provides a method ofinhibiting mutated EGFR in a cell comprising contacting the cell with aneffective amount of at least one compound of Formulae (I)-(VIII) or asalt thereof, and/or with at least one pharmaceutical composition of theembodiments, wherein the contacting is in vitro, ex vivo, or in vivo. Insome embodiments, the mutated EGFR comprises a T790M mutation.

In the inhibitory methods of the embodiments, an “effective amount”means an amount sufficient to inhibit mutated EGFR but not the wild typeEGFR. In some embodiments, the mutated EGFR comprises a T790M mutation.Measuring the degree of inhibition may be performed by routineanalytical methods such as those described below. Such modulation isuseful in a variety of settings, including in vitro assays. Othersettings include ex vivo and in vivo.

In treatment methods according to the embodiments, an “effective amount”means an amount or dose sufficient to generally bring about the desiredtherapeutic benefit in subjects needing such treatment. Effectiveamounts or doses of the compounds of the embodiments may be ascertainedby routine methods, such as modeling, dose escalation, or clinicaltrials, taking into account routine factors, e.g., the mode or route ofadministration or drug delivery, the pharmacokinetics of the agent, theseverity and course of the infection, the subject's health status,condition, and weight, and the judgment of the treating physician. Anexemplary dose is in the range of about 1 μg to 2 mg of active agent perkilogram of subject's body weight per day, preferably about 0.05 to 100mg/kg/day, or about 1 to 35 mg/kg/day, or about 0.1 to 10 mg/kg/day. Thetotal dosage may be given in single or divided dosage units (e.g., BID,TID, QID).

Once improvement of the patient's disease has occurred, the dose may beadjusted for preventative or maintenance treatment. For example, thedosage or the frequency of administration, or both, may be reduced as afunction of the symptoms, to a level at which the desired therapeutic orprophylactic effect is maintained. Of course, if symptoms have beenalleviated to an appropriate level, treatment may cease. Patients may,however, require intermittent treatment on a long-term basis upon anyrecurrence of symptoms. Patients may also require chronic treatment on along-term basis.

Drug Combinations

The methods of the embodiments comprise administering an effectiveamount of at least one compound of Formula (I)-(VIII) or the embodimentsthereof; optionally the compound may be administered in combination withone or more additional therapeutic agents, particularly therapeuticagents known to be useful for treating a proliferative disorder orcancer afflicting the subject.

The additional active ingredients may be administered in a separatepharmaceutical composition from a compound of the embodiments or may beincluded with a compound of the embodiments in a single pharmaceuticalcomposition. The additional active ingredients may be administeredsimultaneously with, prior to, or after administration of a compound ofthe embodiments.

Chemical Synthesis

Exemplary chemical entities useful in methods of the embodiments willnow be described by reference to illustrative synthetic schemes fortheir general preparation below and the specific examples that follow.Artisans will recognize that, to obtain the various compounds herein,starting materials may be suitably selected so that the ultimatelydesired substituents will be carried through the reaction scheme with orwithout protection as appropriate to yield the desired product.Alternatively, it may be necessary or desirable to employ, in the placeof the ultimately desired substituent, a suitable group that may becarried through the reaction scheme and replaced as appropriate with thedesired substituent. Furthermore, one of skill in the art will recognizethat the transformations shown in the schemes below may be performed inany order that is compatible with the functionality of the particularpendant groups. Each of the reactions depicted in the general schemes ispreferably run at a temperature from about 0° C. to the refluxtemperature of the organic solvent used. Unless otherwise specified, thevariables are as defined above in reference to Formula (I). One ofordinary skill in the art will also recognize that the methods describedin these exemplary schemes are also applicable to the preparation ofcompounds of Formula (VIII), as well as compounds of Formulae(II)-(VII).

A representative synthesis for subject compounds is shown in Scheme 1.

In Scheme 1, the variables are as defined herein. As discussed below,X^(2a) and X^(2b) comprise a leaving group. Starting materials may beobtained from commercial sources or via well-established syntheticprocedures.

Referring to Scheme 1, reaction of Compound 1-A with Compound 1-Bthrough a nucleophilic reaction forms Compound 1-C. In Compound 1-A, thehydroxyl group is a nucleophile that can provide the ether linkage inCompound 1-C. The nucleophile can react in a nucleophilic substitutionin which the nucleophile displaces a leaving group on the otherreactant. In alternative embodiments, aniline or thiophenol analogs of1-A are used to access compounds in which X¹ is NH or S. In Compound1-B, X^(2a) comprises a leaving group. Examples of leaving groupsinclude, but are not limited to, halo, triflate, fluorosulfonate,tosylate, or mesylate.

With continued reference to Scheme 1, reaction of Compound 1-C withCompound 1-D under conditions of Buchwald-Hartwig cross-couplingreaction provides Compound 1-E. In Compound 1-D, the amino group is anucleophile that can provide the amino linkage in Compound 1-E. Thenucleophile can react in a nucleophilic aromatic substitution in whichthe nucleophile displaces a leaving group on the other reactant. InCompound 1-C, X^(2b) comprises a leaving group. Examples of leavinggroups include, but are not limited to, halo, triflate, fluorosulfonate,tosylate, or mesylate.

With continued reference to Scheme 1, the nitro group in Compound 1-E isreduced to give an amino group in Compound 1-F. The reduction of nitrogroup can be carried out using an acid catalyst and a metal, or using ametallic catalyst under hydrogen gas. In the acid catalyst reaction,iron, zinc, lithium, sodium, or tin (typically, tin chloride) can beused as the metal, and inorganic acids such as hydrochloric acid,sulfuric acid, nitric acid, or phosphoric acid; organic carboxylic acidssuch as acetic acid or trifluoroacetic acid; amine acid salts such asammonium chloride, can be used as the acid catalyst. Also, in thereduction using a metallic catalyst under hydrogen gas, palladium,nickel, platinum, ruthenium, or rhodium, can be used as the metalliccatalyst.

With continued reference to Scheme 1, amidation of Compound 1-F gives acompound of Formula (I). In the amidation reaction, Compound 1-F reactswith an acryloyl derivative comprising a leaving group. Examples ofleaving groups include, but are not limited to, halo, triflate,fluorosulfonate, tosylate, or mesylate. An amidation reaction can becarried out in a solvent, such as dimethylformamide or dichloromethane,in the presence of a base, such as triethylamine ordiisopropylethylamine. The amidation reaction can be conducted using acoupling agent, such as for example, dicyclohexylcarbodiimide (DCC),1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), orN-[dimethylamino-1H-1,2,3-triazole[4,5-b]-pyridin-1-ylmethylene]-N-methyl-methaneaminium(HATU) together with 1-hydroxy-1H-benzotriazole (HOBT).

In certain embodiments, a representative synthesis for subject compoundsis shown in Scheme 2.

In certain embodiments, a representative synthesis for subject compoundsis shown in Scheme 3.

In certain embodiments, a representative synthesis for subject compoundsis shown in Scheme 4.

In certain embodiments, a representative synthesis for subject compoundsis shown in Scheme 5.

Compounds in which X¹ is NH are prepared according to Scheme 5-1, asshown for exemplary Compound 35.

Accordingly and as described in more detail herein, the presentdisclosure provides a process of preparing a compound of the presentdisclosure, the process involves:

reacting a compound of formula

thereby producing a compound of formula

wherein R³, R⁶, R⁷, R⁸, R¹⁰, R¹¹, R¹², R¹³, Q, and n are defined hereinand X² is a leaving group.

Accordingly and as described in more detail herein, the presentdisclosure provides a process of preparing a compound of the presentdisclosure, the process involves:

reacting a compound of formula

thereby producing a compound of formula

wherein X¹, R³, R⁶, R⁷, R⁸, R¹⁰, R¹¹, R¹², R¹³, R¹⁸, R¹⁹, Q, and n aredefined herein and X² is a leaving group.

Accordingly and as described in more detail herein, the presentdisclosure provides a process of preparing a compound of the presentdisclosure, the process involves:

reducing the nitro group of the compound of formula

and

performing an amidation reaction with an acryloyl derivative comprisinga leaving group;

thereby producing a compound of Formula (I).

Accordingly and as described in more detail herein, the presentdisclosure provides a process of preparing a compound of the presentdisclosure, the process involves:

reducing the nitro group of the compound of formula

and

performing an amidation reaction with an acryloyl derivative comprisinga leaving group;

thereby producing a compound of Formula (VIII).

In certain instances, the above processes further involving the step offorming a salt of a compound of the present disclosure. Embodiments aredirected to the other processes described herein; and to the productprepared by any of the processes described herein.

EXAMPLES

The following examples are offered to illustrate but not to limit theinvention.

Example 1 Synthesis of Compounds 1 and 4

A synthesis ofN-(3-((7-(hydroxymethyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide(Compound 4) andN-(3-((2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamide(Compound 1) and their intermediates is shown in Scheme 6 and describedbelow.

Synthesis of2,4-dichloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(Compound 6-B)

Sodium hydride (60%, 46.7 mg, 3.06 mmol) was added to a mixture ofCompound 2-A (575 mg, 3.06 mmol) and 2-(trimethylsilyl)ethoxymethylchloride (561 mg, 3.37 mmol) in tetrahydrofuran (5 mL) at 0° C. withstirring. The reaction mixture was allowed to warm to room temperatureand stirred for 3 hours before quenching with water (5 mL). The mixturewas extracted with ethyl acetate (10 mL×3). The organic layers werecombined, washed with brine, dried over Na₂SO₄ and filtered. Thefiltrate was concentrated, and the crude material was purified by columnchromatography (PE/EA=20/1) to give Compound 6-B (520 mg, yield 53.4%,M+H⁺=319.27) as a pale yellow solid.

Synthesis of2-chloro-4-(3-nitrophenoxy)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(Compound 6-C)

To a mixture of Compound 6-B (200 mg, 0.628 mmol) and 3-nitrophenol(96.2 g, 0.691 mmol) in dimethylformamide (2 mL) was added K₂CO₃ (173.7mg, 1.26 mmol). The reaction mixture was stirred at room temperature for4 hours. The reaction mixture was then filtered. The filtrate wasdiluted with water, then extracted with ethyl acetate. The organic layerwas washed with water, brine, and dried over Na₂SO₄. After filtrationand removal of the volatiles in vacuo, the crude product was purified byflash column chromatography (PE/EA=20/1) affording Compound 6-C (200 mg,yield 75.6%, M+H⁺=421.92) as a white solid.

Synthesis of2-chloro-6-(difluoro(3-nitrophenyl)methyl)-9-(tetrahydro-2H-pyran-2-yl)-9H-purine(Compound 6-D)

A mixture of Compound 6-C (150 mg, 0.356 mmol), 4-(4-methylpiperazino)aniline (70 mg, 0.356 mmol), tris(dibenzylideneacetone)dipalladium (36mg, 0.0356 mmol), dicyclohexyl (2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (100 mg, 0.214 mmol) and potassium carbonate (197 mg, 1.424mmol) in tert-butanol (8 mL) was stirred under argon at 80° C.overnight. After cooling to room temperature, the reaction mixture wasfiltered through a pad of Celite. The pad of Celite was washed withmethanol and the filtrate was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (DCM/MeOH=20/1) togive Compound 6-D (180 mg, M+H⁺=576.23) as yellow solid.

Synthesis of4-(3-aminophenoxy)-N-(4-(4-methylpiperazin-1-yl)phenyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound

Compound 6-D (180 mg, 0.312 mmol) was dissolved in ethanol (6 mL) andwater (2 mL) was added. Iron powder (90 mg, 1.61 mmol) and ammoniumchloride (230 mg, 4.3 mmol) were then added, and the resulting mixturewas heated at reflux for 3 hours. The reaction mixture was cooled toroom temperature and filtered through a pad of Celite. The ethanol wasremoved in vacuo, and the resulting residue was basified with sodiumbicarbonate and extracted with ethyl acetate. The organic layer wasseparated and dried using anhydrous sodium sulfate, concentrated, andpurified by flash chromatography with 20:1 dichloromethane-methanol toafford Compound 6-E (170 mg, M+H⁺=546) as a white solid.

Synthesis ofN-(3-(2-(4-(4-methylpiperazin-1-yl)phenylamino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 6-F)

Acryloyl chloride (33.8 mg, 0.374 mmol) was added dropwise to a solutionof Compound 6-E (170 mg, 0.312 mmol) and diisopropyethylamine (55 mg,0.426 mmol) in methylene chloride (3 mL) at 0° C. The reaction mixturewas stirred for 1 hour. Water was added to quench the reaction. Theorganic layer was washed with water, brine, and dried over Na₂SO₄. Afterfiltration, removal of volatiles was performed in vacuo. The crudeproduct was purified by flash chromatography (DCM/MeOH=20/1) affordCompound 6-F (125 mg, yield 66.9%, M+H⁺=600.8) as a white solid.

Synthesis ofN-(3-(7-(hydroxymethyl)-2-(4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 4)

Compound 6-F (125 mg, 0.208 mmol) in methylene chloride (3 mL) andtrifluoroacetic acid (1 mL) was stirred at room temperature for 3 hours.Monitoring by thin layer chromatography indicated that all startingmaterial had been consumed. Saturated aqueous NaHCO₃ was then added tothe reaction mixture at 0° C. The reaction mixture was extracted withmethylene chloride. The organic layer was washed with water, brine,dried over Na₂SO₄ and filtered. The filtrate was concentrated and thecrude material was purified by column chromatography (DCM/MeOH=20/1) togive Compound 4 (70 mg, yield 71.5%, M+H⁺=500.5) as a white solid.

Synthesis ofN-(3-(2-(4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 1)

A solution of Compound 4 (100 mg, 0.2 mmol) in methanol (2 mL) wassaturated with ammonia. The reaction mixture was stirred overnight atroom temperature. Monitoring by LC-MS indicated that all startingmaterial had been consumed. The solvent was concentrated and the crudematerial was purified by column chromatography (DCM/MeOH=20/1) to giveCompound 1 (60 mg, yield 63.8%, M+H⁺=470.5) as a pale yellow solid.

Example 2 Synthesis of Compounds 2 and 5

A synthesis ofN-(3-(7-(hydroxymethyl)-2-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 5) andN-(3-(2-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 2) and their intermediates is shown in Scheme 7 and describedbelow.

Synthesis ofN-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-4-(3-nitrophenoxy)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound 7-B)

Compound 7-B (yield 62% from 3, M+H⁺=577.3) was prepared according tothe procedure of Compound 6-D using3-amino-6-(4-Methyl-1-piperazinyl)pyridine hydrochloride instead of4-(4-Methylpiperazino) aniline.

Synthesis of4-(3-aminophenoxy)-N-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound

Compound 7-C(yield 80% from Compound 7-B, M+H⁺=547.3) was preparedaccording to the procedure of Compound 6-E.

Synthesis ofN-(3-(2-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 7-D)

Compound 7-D (yield 67% from Compound 7-C, M+H⁺=601.3) was preparedaccording to the procedure of Compound 6-F.

Synthesis ofN-(3-(7-(hydroxymethyl)-2-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 5)

Compound 7-E (yield 70% from Compound 5, M+H⁺=501.6) was preparedaccording to the procedure of Compound 4.

Synthesis ofN-(3-(2-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 2)

Compound 2 (yield 62% from Compound 5, M+H⁺=471.5) was preparedaccording to the procedure of Compound 1.

Example 3 Synthesis of Compounds 3 and 6

A synthesis ofN-(3-(2-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)-7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 6) andN-(3-(2-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 3) and their intermediates is shown in Scheme 8 and describedbelow.

Synthesis ofN-(3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)-4-(3-nitrophenoxy)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound 8-B)

Compound 8-B (yield % from Compound 8-A, M+H⁺=594.3) was preparedaccording to the procedure of Compound 6-D using3-fluoro-4-(4-methylpiperazin-1-yl)aniline instead of4-(4-methylpiperazino) aniline.

Synthesis of4-(3-aminophenoxy)-N-(3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound 8-C)

Compound 8-C (yield 85% from Compound 8-B, M+H⁺=564.3) was preparedaccording to the procedure of Compound 6-E.

Synthesis ofN-(3-(2-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 8-D)

Compound 8-D (yield 75% from Compound 8-C, M+H⁺=618.3) was preparedaccording to the procedure of Compound 6-F.

Synthesis ofN-(3-(2-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)-7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 6)

Compound 6 (yield 78% from Compound 8-D, M+H⁺=518.6) was preparedaccording to the procedure of Compound 4.

Synthesis ofN-(3-(2-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 3)

Compound 3 (yield 83% from Compound 6, M+H⁺=488.5) was preparedaccording to the procedure of Compound 1.

Example 4N-(3-(7-(2-fluoroethyl)-2-(4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 7)

A synthesis ofN-(3-(7-(2-fluoroethyl)-2-(4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 7) and its intermediates is shown in Scheme 9 and describedbelow.

Synthesis of 2,4-dichloro-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidine(Compound 9-B)

Sodium hydride (60%, 424 mg, 10.6 mmol) was added to a mixture ofCompound 9-A (1 g, 5.3 mmol) and BrCH₂CH₂F (1.519 g, 11.9 mmol) inacetonitrile (10 mL) at room temperature. The reaction mixture wasstirred for 4 hours before quenching with water and then extracted withethyl acetate. The organic layer was washed with brine, dried overNa₂SO₄ and filtered. The filtrate was concentrated and the crudematerial was purified by column chromatography (PE/EA=20/1) to giveCompound 9-B (1.1 g, yield 90%, M+H⁺=234.0) as a pale yellow solid.

Synthesis of2-chloro-7-(2-fluoroethyl)-4-(3-nitrophenoxy)-7H-pyrrolo[2,3-d]pyrimidine(Compound 9-C)

Compound 9-C (yield 82% from Compound 9-B, M+H⁺=337.0) was preparedaccording to the procedure of Compound 6-C.

Synthesis of7-(2-fluoroethyl)-N-(4-(4-methylpiperazin-1-yl)phenyl)-4-(3-nitrophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound 9-D)

Compound 9-D (yield 73% from Compound 9-C, M+H⁺=492.2) was preparedaccording to the procedure of Compound 6-D.

Synthesis of4-(3-aminophenoxy)-7-(2-fluoroethyl)-N-(4-(4-methylpiperazin-1-yl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(Compound 9-E)

Compound 9-E (yield 81% from Compound 9-D, M+H⁺=462.2) was preparedaccording to the procedure of Compound 6-E.

Synthesis ofN-(3-(7-(2-fluoroethyl)-2-(4-(4-methylpiperazin-1-yl)phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 7)

Compound 7 (yield 77% from Compound 9-E, M+H⁺=516.6) was preparedaccording to the procedure of Compound 6-F.

Example 5 Synthesis ofN-(3-(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 34)

Synthesis of tert-butyl3-(3-methoxy-4-nitrophenylamino)azetidine-1-carboxylate

Into a 100 mL 3-Neck round-bottomed flask equipped with reflux condenserwere charged 4-fluoro-2-methoxy-1-nitrobenzene (4.086 g) and tert-butyl3-aminoazetidine-1-carboxylate (4.4 g), triethylamine (9.6 mL), anddimethyl sulfoxide (20 mL). The reaction mixture was heated at 95° C.for 8 hours. The reaction mixture was poured into water (200 mL) andextracted with ethyl acetate (50 mL×3). The organic layer was washedwith brine (50 mL×2), dried over sodium sulfate, and concentratedcompletely under reduced pressure at 40° C. to give the title compound(9 g) which was used without further purification.

Synthesis of N-(3-methoxy-4-nitrophenyl)azetidin-3-amine

To tert-butyl 3-(3-methoxy-4-nitrophenylamino)azetidine-1-carboxylate (9g) was added TFA (18 mL) at room temperature. The reaction mixture wasstirred for 15 min at room temperature and then was concentrated underreduced pressure at 40° C. to give the title compound as TFA salt (7.24g).

Synthesis of1-(2-fluoroethyl)-N-(3-methoxy-4-nitrophenyl)azetidin-3-amine

To N-(3-methoxy-4-nitrophenyl)azetidin-3-amine (3 g) were added Cs₂CO₃(12 g) and 1,2-bromofluoroethane (1.5 g) in DMF (30 mL). The reactionmixture was heated at 50° C. for 8 h. The reaction mixture was poured inwater and extracted in ethyl acetate (100 mL×3). The organic layer waswashed with brine (100 mL x2), dried over sodium sulfate, andconcentrated under reduced pressure. The crude material was purified bycolumn chromatography (DCM/MeOH=50/1 as elution) to give the titlecompound (1.35 g, yield 51% over 3 steps) as a yellow solid.

Synthesis ofN1-(1-(2-fluoroethyl)azetidin-3-yl)-3-methoxybenzene-1,4-diamine

A solution of1-(2-fluoroethyl)-N-(3-methoxy-4-nitrophenyl)azetidin-3-amine (2.6 g)and Pd/C (1 g) in 1,4-dioxane (50 mL) was hydrogenated for 4 hours atroom temperature. The reaction mixture was filtered through diatomaceousearth, washing with MeOH. The filtrate was concentrated and purified bycolumn chromatography (DCM/MeOH=50/1 as elution) to provide the titlecompound (1.57 g, yield 68%, M+H⁺=240.2).

Synthesis of(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-4-(3-nitrophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate

A mixture ofN1-(1-(2-fluoroethyl)azetidin-3-yl)-3-methoxybenzene-1,4-diamine (870mg, 3.64 mmol) and(2-chloro-4-(3-nitrophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (1.55 g, 3.83 mmol), potassium carbonate (1.35 g, 9.77 mmol),tris(dibenzylideneacetone)dipalladium (173 mg, 0.19 mmol) anddicyclohexyl (2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (222 mg, 0.47mmol), a magnetite, and t-BuOH (35 mL) was heated to reflux and stirredunder nitrogen for 2 h. The mixture was cooled to 40-50° C. and wasfiltered through diatomaceous earth, washing with ethyl acetate (50 mL).The filtrate was concentrated under reduced pressure. The crude materialwas purified by column chromatography (DCM/MeOH=50/1 as elution) to givethe title compound (1.7 g, yield 74%, M+H⁺=608.3) as a light yellowsolid.

Synthesis ofN1-(4-(3-aminophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N4-(1-(2-fluoroethyl)azetidin-3-yl)-2-methoxybenzene-1,4-diamine

A mixture of(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-4-(3-nitrophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (530 mg, 0.87 mmol), NH₂NH₂.H₂O (98%, 2.5 mL), Pd/C (110 mg), amagnetite, and MeOH (10 mL) was stirred at reflux temperature overnight.The mixture was cooled to room temperature, and was filtered throughdiatomaceous earth, washing with MeOH (20 mL). The filtrate wasconcentrated under reduced pressure. NaHCO_(3(aq)) was added, and themixture was extracted with ethyl acetate (30 mL×3). The combined organiclayers were concentrated under reduced pressure. The crude material waspurified by column chromatography (DCM/MeOH=40/1 as elution) to give thetitle compound (125 mg, yield 31%, M+H⁺=464.2) as a white solid.

Synthesis ofN-(3-(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenyl)acrylamide(Compound 34)

A 50 mL-round-bottom flask with a magnetite was charged withN1-(4-(3-aminophenoxy)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N4-(1-(2-fluoroethyl)azetidin-3-yl)-2-methoxybenzene-1,4-diamine(125 mg, 0.27 mmol), diisopropylethylamine (43 mg, 0.33 mmol) and DCM(20 mL). The mixture was cooled with an ice bath until the temperaturewas below 0° C., and a solution of acryloyl chloride (33 mg, 0.33 mmol)in THF (2 mL) was added dropwise over 5 minutes. The title compound isisolated and purified by preparative HPLC or preparative LC/MS, or byother standard purification techniques.

Additional exemplary compounds not shown in these synthetic examples areprepared from appropriate starting materials using methods analogous tothose described in the preceding schemes and examples.

Biological Example A In Vitro Cell-Based Screening Using Real-Time CellElectronic Sensing (RT-CES) System

Some assays and examples demonstrating the anti-cancer effects of thecompounds of the embodiments are described as below.

The pyrrolopyrimidine compounds in the embodiments are developed for theanticancer activities for cancer cells with certain molecular targets,i.e., EGFR (epidermal growth factor receptor). The anticancer efficacyof the pyrrolopyrimidine compounds may be preliminarily screened invitro using a panel of EGFR cancer cell lines by real time electroniccell sensing (RT-CES) system from ACEA Biosciences, Inc. (or xCELLigencesystem from Roche Applied Sciences/ACEA Biosciences Inc.), whichprovides dynamic cell response information after exposing to ananticancer agent.

The details of this cell electronic sensing technology, called real-timecell electronic sensing (RT-CES®) and associated devices, systems andmethods of use are described in U.S. Pat. No. 7,732,127; U.S. Pat. No.7,192,752; U.S. Pat. No. 7,459,303; U.S. Pat. No. 7,468,255; U.S. Pat.No. 7,470,533; U.S. Pat. No. 7,560,269; U.S. provisional application No.60/397,749, filed on Jul. 20, 2002; U.S. provisional application No.60/435,400, filed on Dec. 20, 2002; U.S. Provisional application60/469,572, filed on May 9, 2003, PCT application number PCT/US03/22557,filed on Jul. 18, 2003; PCT application number PCT/US03/22537, filed onJul. 18, 2003; PCT application number PCT/US04/37696, filed on Nov. 12,2004; PCT application number PCT/US05/04481, filed on Feb. 9, 2005; U.S.patent application Ser. No. 10/705,447, filed on Nov. 10, 2003; U.S.patent application Ser. No. 10/705,615, filed on Nov. 10, 2003; U.S.patent application Ser. No. 10/987,732, filed on Nov. 12, 2004; U.S.patent application Ser. No. 11/055,639, filed on Feb. 9, 2005, each ofwhich is incorporated by reference. Additional details of RT-CEStechnology is further disclosed in U.S. provisional application No.60/519,567, filed on Nov. 12, 2003, and U.S. provisional application No.60/542,927, filed on Feb. 9, 2004, U.S. provisional application No.60/548,713, filed on Feb. 27, 2004, U.S. provisional application No.60/598,608, filed on Aug. 4, 2004; U.S. provisional application No.60/598,609, filed on Aug. 4, 2004; U.S. provisional application No.60/613,749, filed on Sep. 27, 2004; U.S. provisional application No.60/613,872, filed on Sep. 27, 2004; U.S. provisional application No.60/614,601, filed on Sep. 29, 2004; U.S. provisional application No.60/630,071, filed on Nov. 22, 2004; U.S. provisional application No.60/630,131, filed on Nov. 22, 2004, each of which is incorporated hereinby reference.

For measurement of cell-substrate or cell-electrode impedance usingRT-CES technology, microelectrodes having appropriate geometries arefabricated onto the bottom surfaces of microtiter plate or similardevice, facing into the wells. Cells are introduced into the wells ofthe devices, and make contact to and attach to the electrode surfaces.The presence, absence or change of properties of cells affects theelectronic and ionic passage on the electrode sensor surfaces. Measuringthe impedance between or among electrodes provides information aboutbiological status of cells present on the sensors. When there arechanges to the biological status of the cells analogue, electronicreadout signals are measured automatically and in real time, and areconverted to digital signals for processing and analysis.

In a RT-CES system, a cell index is automatically derived and providedbased on measured electrode impedance values. The cell index obtainedfor a given well reflects: 1) how many cells are attached to theelectrode surfaces in this well; and 2) how well cells are attached tothe electrode surfaces in this well. Thus, the more the cells of sametype in similar physiological conditions attach the electrode surfaces,the larger the cell index. And, the better the cells attach to theelectrode surfaces (e.g., the cells spread-out more to have largercontact areas, or the cells attach tighter to electrode surfaces), thelarger the cell index. We have found that the cMet-addictive cell lineswould produce a transient impedance response profile when treated withpositive-control EGFR (epidermal growth factor receptor) inhibitors.

Through the use of the RT-CES system, the pyrrolopyrimidine compoundsdescribed in the examples above have been shown to produce a similarcell response impedance profile on RT-CES system to that generated bypositive control inhibitors. In addition, these compounds have beenshown to inhibit EGFR (epidermal growth factor receptor)-induced cellmigration in several cell lines. In addition, these compounds have shownno or negligible effects when they were used to treat non-cMet addictivecancer cell lines.

The RT-CES system (or xCELLigence RTCA system) comprises threecomponents, an electronic sensor analyzer, a device station and 16× or96× microtiter plate devices (i.e. E-Plate 16 or E-Plate 96).Microelectrode sensor array was fabricated on glass slides withlithographical microfabrication methods and the electrode-containingslides are assembled to plastic trays to form electrode-containingwells. Each 16× (or 96×) microtiter plate device used in RT-CES systemcomprises up to 16 (or 96) such electrode-containing wells. The devicestation receives the 16× or 96× microtiter plate devices and is capableof electronically switching any one of the wells to the sensor analyzerfor impedance measurement. In operation, the devices with cells culturedin the wells are placed into a device station (xCELLigence RTCA SPstation or RT-CES SP station) that is located inside an incubator.Electrical cables connect the device station to the sensor analyzer(xCELLigence RTCA analyzer or RT-CES analyzer). Under the RT-CES orxCELLigence RTCA software control, the sensor analyzer can automaticallyselect wells to be measured and continuously conduct impedancemeasurements. The impedance data from the analyzer is transferred to acomputer, analyzed and processed by the integrated software.

Impedance measured between electrodes in an individual well depends onelectrode geometry, ionic concentration in the well and whether thereare cells attached to the electrodes. In the absence of the cells,electrode impedance is mainly determined by the ion environment both atthe electrode/solution interface and in the bulk solution. In thepresence of the cells, cells attached to the electrode sensor surfaceswill alter the local ionic environment at the electrode/solutioninterface, leading to an increase in the impedance. The more cells thereare on the electrodes, the larger the increase in cell-electrodeimpedance. Furthermore, the impedance change also depends on cellmorphology and the extent to which cells attach to the electrodes.

To quantify cell status based on the measured cell-electrode impedance,a parameter termed Cell Index is derived, according to

${CI} = {\max\limits_{{i = 1},\; \ldots \mspace{11mu},N}\left( {\frac{R_{cell}\left( f_{i} \right)}{R_{b}\left( f_{i} \right)} - 1} \right)}$

where R_(b)(f) and R_(cell)(f) are the frequency dependent electroderesistances (a component of impedance) without cells or with cellpresent, respectively. N is the number of the frequency points at whichthe impedance is measured. Thus, Cell Index is a quantitative measure ofthe status of the cells in an electrode-containing well. Under the samephysiological conditions, more cells attached on to the electrodes leadsto larger R_(cell)(f) value, leading to a larger value for Cell Index.Furthermore, for the same number of cells present in the well, a changein the cell status such as morphology will lead to a change in the CellIndex. For example, an increase in cell adhesion or cell spreading leadsto larger cell-electrode contact area which will lead to an increase inR_(cell)(f) and thus a larger value for Cell Index. The Cell Index mayalso be calculated using a formula different from the one describedhere. Other methods for calculating the Cell Index based on impedancemeasurement can be found in U.S. Pat. No. 7,732,127; U.S. Pat. No.7,192,752; U.S. Pat. No. 7,459,303; U.S. Pat. No. 7,468,255; U.S. Pat.No. 7,470,533; U.S. Pat. No. 7,560,269; PCT application numberPCT/US04/37696, fined on Nov. 12, 2004, PCT application numberPCT/US05/04481, filed on Feb. 9, 2005, U.S. patent application Ser. No.10/987,732, filed on Nov. 12, 2004, and U.S. patent application Ser. No.11/055,639, filed on Feb. 9, 2005.

Biological Example B-1 Bioactivity of Pyrrolopyrimidine Compounds onEGFR Mutated Cell Lines Material and Methods

Cell Culture and Reagents

All cell lines were obtained from the American Type Culture Collectionand were maintained at 37° C. with 5% CO₂, in media supplemented with10% fetal bovine serum and 1% L-glutamine-penicillin-streptomycin. H1975and HCC827 cells were cultured with RPMI 1640 media. A431 cells weremaintained in Dulbecco's Modification of Eagle's Medium EGF (R&D), EGFinhibitors were resuspended and stored according to the manufacturers'instructions.

Cell Proliferation and Growth Inhibition Assay

Cell proliferation was assessed by WST assay (Roche, Indianapolis, Ind.)per the manufacturer's instructions. The H1975, HCC827 and A431 cellswere seeded at 3,000, 3,000 and 4,000 cells per well onto 96-wellplates, and after a 24-hour incubation, the cells were treated with testcompounds for 72 hours. Cell viability was assayed by incubating thecells with WST-1 reagent for 2 hours, and then with the measurement ofthe absorbance at a wavelength of 450 nm. The data was calculated usingGraphPad Prism version 4.0. The IC₅₀ values were fitted using anon-linear regression model with a sigmoidal dose response.

Western Blotting

H1975 and A431 cells were seeded onto 6-well plates at a concentrationof 1×10⁶ cells per well. After 24 hours of growth in serum-containingmedia, cells were incubated in serum-free media for 1 hour, and thentreated with test compound for 2 hour. A431 cells were stimulated with30 ng/mL EGF during the last 20 minutes of compound treatment. Westernblots were done on the whole-cell extracts using phospho-specific EGFR(pY1068), total EGFR, phospho-Akt (Ser-473), total Akt, phospho-ERK1/2(pT202/pY204) and total ERK1/2 antibodies(Cell Signaling Technology).

Tumor sections were snap-frozen in liquid nitrogen for proteinisolation, and EGFR signal transduction was evaluated by Western blotwith primary antibodies included the following: phospho-specific EGFR(pY1068), total EGFR, phospho-Akt (Ser-473), total Akt, phospho-ERK1/2(pT202/pY204) and total ERK1/2.

ELISA Assay

H1975 and A431 cells were seeded onto each well of a 96-well plate at adensity of 4×10⁴ cells per well. After 24 hours of growth inserum-containing media, cells were treated with test compound inserum-free medium for 2 hours. A431 cells were stimulated with 30 ng/mLEGF during the last 15 minutes of compound treatment. Cells were washedwith ice cold PBS before extraction with 100 n1 per well cell lysisbuffer. Phosphorylation of EGFR was measured using a sandwich ELISAassay with the pair of phospho-specific EGFR (pY1068) and total EGFRantibodies.

Results

Compound 3 Inhibits Proliferation of EGFR− Mutant Cells

The following compounds were tested.

Sensitivity of cancer cell lines that express EGFR WT, Exon 19 Del,L858R/T790M and delE746-A750 to Compound 3, Compound B and gefitinib.Cell proliferation assays were performed with increasing concentrationsof compounds for 72 hours using WST. IC₅₀ values were determined byGraphPad software. Compound 3 inhibits proliferation of the T790M−positive H1975 cells more potently than gefitinib.

TABLE 1 H1975 cell A431 cell HCC827 cell Compound (T790M/L858R) (WT)(delE746-A750) Compound 3 0.61 μM 10.8 μM 0.019 μM Compound B  1.1 μM 4.5 μM 0.013 μM Gefitinib  >10 μM ND 0.024 μM

Compound 3 Inhibits EGFR Phosphorylation in H1975 Cells

Inhibition of EGFR phosphorylation and proliferation in H1975 cellstreated with Compound 3. H1975 and A431 cells were incubated withvarious concentrations of Compound 3 or Compound B for 2 hours, and thewhole cell extracts were directly harvested and tested for pEGFR byELISA. IC₅₀ values were determined by GraphPad software.

TABLE 2 A431 cell Compound H1975 cell (T790M/L858R) (WT) Compound 30.031 μM 12.7 μM Compound B 0.063 μM  8.9 μM

Compound 3 Inhibits the EGFR Signaling Pathway in H1975 Cells

Exponentially growing H1975 lung cancer cells were treated with Compound3 at indicated concentrations for 2 hours in serum-free medium. As shownin FIG. 1, whole cell extracts were resolved by SDS-PAGE before blottingonto nitrocellular membranes. Inhibition of phosphorylation of EGFRleads to inhibition of its downstream effectors p-Akt and p-ERK. Allantibodies were obtained from Cell Signaling.

Compound 3 Inhibits the EGFR Signaling Pathway in H1975 Tumors

Compound 3 was administered PO at 100 mg/kg, and tumors were harvestedat 1, 4, 8, 18 and 25 hours after the single dose. As shown in FIG. 2,immunoblots were probed for pEGFR, total EGFR, pAkt, total Akt, p-ERKand total ERK. Compound 3 inhibited the phosphorylation of EGFR in atime-dependent manner, and the inhibition at EGFR leads to theinhibition of its downstream effectors p-Akt and p-ERK.

Comparison between Compound 3 and Compound A

1. WST Result

TABLE 3 H1975 cell A431 cell HCC827 cell Compound (T790M/L858R) (WT)(delE746-A750) Compound 3 0.73 μM 0.62 μM 0.011 μM Compounds A 1.63 μM4.17 μM 0.023 μM

2. ELISA Result

TABLE 4 H1975 cell H1975 cell A431 cell (T790M/L858R) Compound(T790M/L858R) (WT) EGF stimulate Compound 3 0.0032 μM 0.4737 μM 0.025 μMCompound A 0.0088 μM 1.0270 μM 0.091 μM

Biological Example B-2 Cell Culture and Reagents

All cell lines were obtained from the American Type Culture Collectionand were maintained at 37° C. with 5% CO2, in media supplemented with10% fetal bovine serum and 1% L-glutamine-penicillin-streptomycin. H1975and HCC827 cells were cultured with RPMI 1640 media. A431 cells weremaintained in Dulbecco's Modification of Eagle's Medium GTL-16 cells,T47D cells and BxPC3 cells were cultured with RPMI 1640 media. NIH-3T3cells, H460 cells and HepG2 cells were cultured with Dulbecco'sModification of Eagle's Medium A549 cells were cultured with F-12KNutrient Mixture media. H295R were cultured with DMEM: F12 Media. WST-1reagent was obtained from Roche. EGF (R&D), EGF inhibitors wereresuspended and stored according to the manufacturers' instructions.

Cell Proliferation and Growth Inhibition Assay

Cell proliferation was assessed by WST assay (Roche, Indianapolis, Ind.)per the manufacturer's instructions. The H1975, HCC-827 and A431 cellswere seeded at 3,000, 3,000 and 4,000 cells per well onto 96-wellplates, and after a 24 h incubation, the cells were treated with testcompounds for 72 hrs. NIH-3T3 cells, A549 cells, H295R cells, GTL-16cells, H460 cells, HepG2 cells, Hela cells, T47D cells and BxPC3 cellswere seeded at 2,000, 2,000, 5,000, 5,000, 2,500, 5,000, 2,000, 5,000and 5,000 cells per well onto 96-well plates. Cell viability was assayedby incubating the cells with WST-1 reagent for 3 hrs. Absorbance wasmeasured at OD450-620 using the Beckman DTX880. The data was calculatedusing GraphPad Prism version 4.0. The IC50 were fitted using anon-linear regression model with a sigmoidal dose response.

ELISA Assays

The H1975, HCC-827 and A431 cells were seeded onto a 96-well plate at adensity of 40,000, 40,000 and 60,000 cells per well respectively. After24 h of growth in serum-containing media, cells were treated with testcompound in serum-free medium for 2 h. A431 cells were stimulated with50 ng/mL EGF during the last 15 min of compound treatment. Cells werewashed with ice cold PBS before extraction with 100 μl per well celllysis buffer. Phosphorylation of EGFR was measured using a sandwichELISA assay with the pair of phospho-specific EGFR (pY1068) and totalEGFR antibodies. The data was calculated using GraphPad Prism version4.0. The IC50 were fitted using a non-linear regression model with asigmoidal dose response.

Western Blotting

H1975, HCC-827 and A431 cells were seeded onto 6-well plates at aconcentration of 1×10⁶ cells per well. After 24 h of growth inserum-containing media, cells were incubated in serum-free media for 1h, and then treated with test compound for 2 h. A431 cells werestimulated with 30 ng/mL EGF during the last 20 min of compoundtreatment. Western blots were performed on the whole-cell extracts usingphospho-specific EGFR (pY1068), total EGFR, phospho-Akt (Ser-473), totalAkt, phospho-ERK1/2 (pT202/pY204) and total ERK1/2 antibodies(CellSignaling Technology). The density of blotting band was acquired usingImageJ software, and the IC50 of EGFR Tyr1068 phosphorylation was fittedusing a non-linear regression model by GraphPad Prism version 4.0.

Compound 3 was orally administered at indicated dose (12.5, 50, 200mg/kg), and Gefitinib (GF) was orally administrated at 100 mg/kg. Thetumor tissues were harvested at 1, 4, 8, and 24 h at Day 1 and after thesingle dose, or harvested at Day 8 and after 8 consecutive doses fordose (12.5, 50 mg/kg). Tumor sections were snap-frozen in liquidnitrogen for protein isolation, and EGFR signal transduction wasevaluated by Western blot with primary antibodies included thefollowing: phospho-specific EGFR (pY1068), total EGFR.

Cell-Based Pulse Chase Assay for Irreversibility Assessment of Compound

The H1975 was seeded at 3000 cell per well in RTCA system (xCELLigenceSP instrument, ACEA Biosciences). After one day culture, cell weretreated with compound of compound 3, WZ4002 at concentration of 10 μMfor 22 hrs then removed compared with drugs kept overtime. About 60hours after recovery, cell subject to WST viability measuring.

Results

Compound 3 inhibited the proliferation of EGFR mutation harboring cancercells.

Compound 3 achieved the inhibition of the proliferation of H1975(T790M/L858R) cells with the IC50 at 91±60 nM, and with IC50 at 19±8 nMfor HCC827(Del E746-A750) cells, whereas the sensitivity to A431(WT)cells is much lower (IC50=2113±1660 nM). In contrast, gefitinib, thefirst generation EGFR inhibitor, exhibited sensitivity to A431 cells,but had no activity on inhibiting the proliferation of T790M mutationharboring cells (IC50>20 uM).

H1975 cell A431 cell HCC827 cell Compound (T790M/L858R) (WT)(delE746-A750) Compound 3 91 ± 60 nM 2113 ± 1660 nM 19 ± 8 nM WZ40021905 ± 732 nM  4393 ± 617 nM 35 ± 12 nM  Gefitinib >20000 nM  523 ± 115nM  9 ± 1 nM

Compound 3 Significantly Reduced the EGFR Tyr1068 Phosphorylation inEGFR Mutant Cells.

H1975 and A431 cells were incubated with various concentrations ofcompound 3 or WZ4002 for 2 h, and the whole cell extracts were directlyharvested and tested for pEGFR by ELISA. IC50 values were determined byGraphPad software.

The cell-based ELISA assays verified that compound 3 significantlyreduced the EGFR Tyr1068 phosphorylation in the EGFR mutant cell lines,while Gefitinib showed inhibition of the phosphorylation to a much lessextent.

H1975 cell A431 cell Compound (T790M/L858R) (WT) compound 3  4 ± 2 nM 650 ± 63 nM WZ4002 32 ± 12 nM 970 ± 340 nM

As shown in the table below, Compound 3 significantly reduced the EGFRTyr1068 phosphorylation and downstream signaling in the EGFR mutantcells, and is less effective in cell line expressing wild-type EGFR.

IC50 (nM) by WB (Tyr1068 phospho) EGFR genotype Cell line compound 3Gefitinib WZ4002 T790M/L858R H1975 4.4 860 21 DelE746-A750 HCC-827 9.85.4 58 Wild Type A431 288 1.6 53 Selectivity (A431/H1975) 65X 0.002X2.5X

As shown in FIGS. 9A and 9B, Compound 3 inhibited EGFR-Tyr1068phosphorylation and downstream signaling in H1975 EGFR mutant cells.Comparative data for Gefitinib and WZ4002 are shown in FIGS. 9C-9F.

As shown in FIGS. 10A and 10B, Compound 3 inhibited EGFR-Tyr1068phosphorylation and downstream signaling in HCC-827 EGFR mutant cells.Comparative data for Gefitinib and WZ4002 are shown in FIGS. 10C-10F.

As shown in FIGS. 11A and 11B, Compound 3 was less effective oninhibiting EGFR-Tyr1068 phosphorylation and downstream signaling in A431cells expressing WT EGFR. Comparative data for Gefitinib and WZ4002 areshown in FIGS. 11C-11F.

Compound 3 Inhibited the Phosphorylation of the EGFR in H1975 Tumors.

Compound 3 significantly inhibited the phosphorylation of the EGFR inH1975 tumor tissues, at all three dosages of 12.5, 50 and 200 mg/kg. Theinhibition of EGFR phosphorylation by Compound 3 was dose- andtime-dependent. In contrast, the inhibition of the phosphorylation ofthe EGFR was not detected for gefitinib with the dosage at 100 mg/kg.

As shown in FIG. 12, Compound 3 inhibits the phosphorylation of the EGFRin H1975 tumor tissues at single-dose of compound 3.

As shown in FIG. 13, Compound 3 inhibits the phosphorylation of the EGFRin H1975 tumor tissues after 8 consecutive doses of compound 3.

As shown in FIG. 14, Compound 3 irreversibly inhibited the proliferationof H1975 cells harboring EGFR T790M mutation. The reversibility ofcompound 3 was assessed by a cell-based pulse chase assay. As shown inFIG. 14, upon compound 3 withdrawal following a 22-hr treatment, theinhibition of the proliferation of H1975 sustained (7.8±1.3%) up to 60hrs. In contrast, the recovery from WZ4002 treatment was at 27±10%. Theresults of this study demonstrated that compound 3 is an irreversibleinhibitor of EGFR, and exhibited strong binding property than WZ4002.

Viability in Comparison with Vehicle Control (%)

compound 3 for WZ4002 for 22 hr treatment 22 hr treatment Viability ofH1975 cells 7.8 ± 1.3 27 ± 10 (%)

1. WST Result

H1975 cell A431 cell HCC827 cell Compound (T790M/L858R) (WT)(delE746-A750) compound 3 0.19 uM 2.03 uM 0.011 uM compound A 1.16 uM9.14 uM 0.023 uM

2. ELISA Result

A431 cell H1975 cell (WT) EGF H1975 cell Compound (T790M/L858R)stimulate (T790M/L858R) compound 3 0.0032 uM 0.4737 uM 0.025 uM compoundA 0.0088 uM 1.0270 uM 0.091 uM

Biological Example C Evaluation of Efficacy of Compound 3 in theTreatment of H1975, HCC827, and A431 Xenograft Mouse Models

This example evaluates the efficacy of Compound 3 in the treatment ofNCI-H1975 (L858R/T790M) human non-small cell lung adenocarcinoma, HCC827(L858R) human lung adenocarcinoma and A431 (WT) human skin epidermoidcarcinoma xenograft tumor models in nude mice. Gefitinib, a firstgeneration of reversible EGFR tyrosine kinase inhibitor, was used as apositive control on those three mouse xenograft tumor models.

Experimental Design and Dosing Schedule

Experimental Design and dosing schedule are shown below.

TABLE 5 NCI-H1975 Model Dosing Dose volume Dosing Days for Group nTreatment (mg/kg) (μl/g) route Solvent dosing Schedule 1 8 Vehicle —16.7 PO PEG system 14 QD 2 8 Compound 3 25 10 PO PEG system 14 QD 3 8Compound 3 50 10 PO PEG system 14 QD 4 8 Compound 3 100 16.7 PO PEGsystem 14 QD 5 8 Gefitinib 100 10 PO 1% tween80 14 QD Note: n: animalnumber; Dosing volume: adjust dosing volume based on body weight; PEGsystem:PEG200:alcohol:5% dextrose = 4:1:5; Treatment schedule wasadjusted if body weight loss >15%.

TABLE 6 HCC827 Model Dosing Dose volume Dosing Days for Group nTreatment (mg/kg) (μl/g) route Solvent dosing Schedule 1 8 Vehicle — 10PO — 35 QD 2 8 Compound 3 50 10 PO PEG system 35 QD 3 8 Compound 3 50 10PO 0.5% MC 35 QD 4 8 Gefitinib 100 10 PO 1% tween80 7 QD

TABLE 7 A431 Model Dosing Dose volume Dosing Days for Group n Treatment(mg/kg) (μl/g) route Solvent dosing Schedule 1 8 Vehicle — 16.7 PO PEGsystem 14 QD 2 8 Compound 3 100 16.7 PO PEG system 14 QD 3 8 Gefitinib100 10 PO 1% tween80 14 QDNote: n: animal number; Dosing volume: adjust dosing volume based onbody weight; PEG system: PEG200: alcohol: 5% dextrose=4:1:5; Treatmentschedule was adjusted if body weight loss>15%.

Animal Housing Animals

Details on the animals are shown below.

-   Species: Mouse-   Strain: Nu/Nu nude-   Age: 7-8 weeks-   Sex: Female-   Body weight: 20-25 g-   Animal supplier: Vital River Laboratories, Beijing, China

Housing Conditions

The mice were kept in Individual Ventilation Cages at constanttemperature and humidity with 4 animals in each cage at ACEA BioscienceHangzhou Inc.

-   -   Temperature: about 20-26° C.    -   Humidity about 40-70%.

IVC Cages: Made of polycarbonate. The size is 300 mm×180 mm×150 mm. Thebedding material is corn cob, which is changed twice per week.

Diet: Animals had free access to irradiation sterilized dry granule foodduring the entire study period.

Water: Animals had free access to sterile drinking water.

Cage identification: The identification labels for each cage containedthe following information: number of animals, sex, strain, datereceived, treatment, study number, group number and the starting date ofthe treatment.

Animal identification: Animals were marked by ear cutting.

Experimental Methods and Procedures Cell Culture

The NCI-H1975, HCC827 and A431 tumor cells were maintained in vitro as amonolayer culture in medium supplemented with 10% fetal bovine serum,100 U/ml penicillin and 100 μg/ml streptomycin at 37° C. in anatmosphere of 5% CO₂ in air as ATCC recommended. The tumor cells wereroutinely subcultured twice weekly by trypsin-EDTA treatment. The cellsgrowing in an exponential growth phase were harvested and counted fortumor inoculation.

Tumor Inoculation

Each mouse was inoculated subcutaneously at the right flank with H1975cells (5×10⁶), HCC827 (5×10⁶) and A431 (5×10⁶) respectively in 0.2 ml ofmedium for tumor development. The treatments were started when the tumorsize reached approximately 200-250 mm³. The testing articles wereadministrated to the mice according to the predetermined regimen asshown in the experimental design table.

Observations

All the procedures related to animal handling, care and the treatment inthis study were performed according to the guidelines approved by theInstitutional Animal Care and Use Committee (IACUC) following theguidance of the Association for Assessment and Accreditation ofLaboratory Animal Care (AAALAC). At the time of routine monitoring, theanimals were checked for any effects of tumor growth and treatments onnormal behavior such as mobility, food and water consumption (byobservation), body weight gain/loss (body weights were measured twiceweekly), eye/hair matting and any other abnormal effect. Death andobserved clinical signs were recorded on the basis of the numbers ofanimals within each subset. Animals that were observed to be in acontinuing deteriorating condition were euthanized prior to death orbefore reaching a comatose state.

Tumor Measurements and the Endpoints

The major endpoint was to see if the tumor growth can be delayed or micecan be cured. Tumor size was measured twice weekly in two dimensionsusing a caliper, and the volume was expressed in mm³ using the formula:V=0.5 a×b² where a and b are the long and short diameters of the tumor,respectively. The tumor size was then used for calculations of both T-Cand T/C values. T-C was calculated with T as the median time (in days)required for the treatment group tumors to reach a predetermined size(e.g., 1,000 mm³), and C as the median time (in days) for the controlgroup tumors to reach the same size. The T/C value (in percent) was anindication of antitumor effectiveness; T and C were the mean volume ofthe treated and control groups, respectively, on a given day. Tumorweight was measured at the study termination. The T/C value (in percent)was calculated where T and C were the mean tumor weights of the treatedand control groups, respectively.

Statistical Analysis

Summary statistics, including mean and the standard error of the mean(SEM), are provided for the tumor volume of each group at each timepoint.

Statistical analysis of difference in tumor volume and tumor weightamong the groups was conducted on the data obtained at the besttherapeutic time point after the final dose (the 15^(th) day after tumorinoculation).

A one-way ANOVA was performed to compare tumor volume and tumor weightamong groups, and when a significant F-statistics (a ratio of treatmentvariance to the error variance) was obtained, comparisons between groupswere carried out with LSD and Games-Howell test. All data were analyzedusing SPSS 16.0. p<0.05 was considered to be statistically significant.

Results 5.1. Body Weights

The results of the body weight changes in the tumor-bearing mice forNCI-H1975, HCC827 and A431 models are shown in FIG. 3, FIG. 4, and FIG.5, respectively.

The mouse body weights in different groups of the tumor-bearing mice atthe end of treatment on NCI-H1975, HCC827 and A431 are shown in Table 8,Table 9 and Table 10 respectively.

TABLE 8 The Mouse Body Weights in the Different Groups on NCI-H1975Model Mouse Weight (g)^(a) Treatment at day 23(14) p Vehicle 22.99 ±0.26 — Compound 3 25 mg/kg po qd 22.28 ± 0.55 0.364 Compound 3 50 mg/kgpo qd 22.73 ± 0.33 0.737 Compound 3 100 mg/kg po qd 22.53 ± 0.66 0.555Gefitinib 100 mg/kg po qd 20.43 ± 0.71 0.002

TABLE 9 The Mouse Body Weights in the Different Groups on HCC827 ModelMouse Weight (g)^(a) Treatment at day 25(14) p Vehicle 23.83 ± 0.71 —Compound 3 (PEG) 50 mg/kg po qd 23.26 ± 0.47 0.523 Compound 3 (MC) 50mg/kg po qd 23.54 ± 0.67 0.743 Gefitinib 100 mg/kg po qd 23.70 ± 0.430.887

TABLE 10 The Mouse Body Weights in the Different Groups on A431 ModelMouse Weight (g)^(a) Treatment at day 25(14) p Vehicle 25.64 ± 0.53 —Compound 3 100 mg/kg po qd 25.66 ± 0.72 0.979 Gefitinib 100 mg/kg po qd21.56 ± 0.51 0.000 Note ^(a)Mean ± SEM

Tumor Volumes

The tumor sizes of the different groups at different time points onNCI-H1975, HCC827 and A431 are shown in Table 11, Table 12, and Table13, respectively.

TABLE 11 Tumor Sizes in the Different Treatment Groups on NCI-H1975model Tumor volume (mm³)^(a) Compound 3, Compound 3, Compound 3,Gefitinib, PO, Vehicle, PO, QD PO, QD PO, QD PO, QD QD Days — 25 mpk 50mpk 100 mpk 100 mpk 9 215.01 ± 20.88 219.91 ± 22.33 215.95 ± 21.58220.64 ± 22.95 215.95 ± 22.36 12 387.98 ± 45.76 284.09 ± 32.64 255.85 ±34.44 181.40 ± 21.15 379.15 ± 46.00 16 828.95 ± 58.76 393.95 ± 42.09268.23 ± 47.77 180.18 ± 26.25 737.84 ± 80.06 19 1425.22 ± 101.9  514.88± 55.57 346.01 ± 62.50 207.28 ± 42.54 1195.5 ± 67.91 23 2169.9 ± 170.8670.36 ± 54.19 373.01 ± 63.35 232.25 ± 37.11 1702.5 ± 101.8

TABLE 12 Tumor Sizes in the Different Treatment Groups on HCC827 modelTumor volume (mm³)^(a) Compound 3, Compound 3, Vehicle, PO, QD PO, QDPO, QD Gefitinib, PO, QD Days Vehicle 50 mpk (PEG) 50 mpk (0.5% MC) 100mpk 14 215.94 ± 25.70 211.90 ± 23.00  211.14 ± 25.11 212.28 ± 26.35  18291.15 ± 24.42 188.72 ± 28.03  216.63 ± 27.69 59.55 ± 25.20 21 353.24 ±25.64 136.96 ± 16.40  245.14 ± 33.44 4.61 ± 3.16 25 453.43 ± 24.72 95.73± 15.38 216.42 ± 28.06 1.25 ± 1.25 28 519.39 ± 22.26 111.96 ± 22.05 231.08 ± 30.81 1.25 ± 1.25 32 638.78 ± 32.70 82.28 ± 24.08 277.59 ±42.02 1.25 ± 1.25 35 762.43 ± 47.22 67.63 ± 24.22 293.64 ± 43.98 1.88 ±1.32 39 1092.53 ± 99.28  69.44 ± 30.35 328.53 ± 43.51 1.88 ± 1.32 421324.76 ± 141.54 79.71 ± 28.86 302.31 ± 35.83 10.95 ± 6.13  46 1736.94 ±217.03 84.26 ± 35.62 284.44 ± 27.00 23.71 ± 11.84 49 1920.11 ± 256.3677.59 ± 42.07 299.28 ± 31.79 41.00 ± 20.52

TABLE 13 Tumor Sizes in the Different Treatment Groups on A431 modelTumor volume (mm³)^(a) Compound 3, Gefitinib, PO, Vehicle, PO, QD PO, QDQD Days — 100 mpk 100 mpk 11 241.34 ± 28.69  240.95 ± 26.46 239.83 ±23.30 14 472.09 ± 71.50  399.68 ± 42.62 203.74 ± 22.97 18 860.82 ±120.62 867.62 ± 70.54 139.70 ± 26.94 21 1211.0 ± 157.77 1166.1 ± 94.08139.70 ± 22.07 25 1666.6 ± 233.36 1627.7 ± 146.0 154.79 ± 32.62 Note^(a)Mean ± SEM

Tumor Growth Inhibition

The tumor growth inhibition on NCI-H1975, HCC827 and A431 models issummarized in Table 14, Table 15, and Table 16, respectively.

TABLE 14 Effect of Compounds in the Treatment of H1975 Xenografts TumorModel T-C Tumor Size (days) (mm³)^(a) T/C at Treatment at day 23(14) (%)300 mm³ p Vehicle 2170 ± 171 — — — Compound 3 25 mg/kg po qd 670 ± 5428.5% 2.11 0.000 Compound 3 50 mg/kg po qd 373 ± 63 16.0% 6.76 0.000Compound 3 100 mg/kg po qd 232 ± 37 9.9% >14 0.000 Gefitinib 100 mg/kgpo qd 1702 ± 102 77.4% 0.08 0.345

TABLE 15 Effect of Compounds in the Treatment of HCC827 Xenografts TumorModel Tumor Size (mm³)^(a) T/C TRR^(b) T-C (days) Treatment at day49(35) (%) at day 49(35) at 300 mm³ P Vehicle 1920 ± 256 — — — —Compound 3 (PEG) 50 mg/kg po qd  78 ± 42 2.8% 71.4% >35 0.001 Compound 3(MC) 50 mg/kg po qd 299 ± 32 14.3% −45.0% 17.3 0.002 Gefitinib 100 mg/kgpo qd  41 ± 21 2.4% 75.7% >35 0.001

TABLE 16 Effect of Compounds in the Treatment of A431 Xenografts TumorModel T-C Tumor Size (days) (mm³)^(a) T/C at Treatment at day 25(14) (%)300 mm³ p Vehicle 1667 ± 233 — — — Compound 3 100 mg/kg po qd 1628 ± 14698.3% 0.35 0.999 Gefitinib 100 mg/kg po qd 154 ± 33 8.7% >14 0.001 Note^(a)Mean ± SEM b. Tumor regression rate (%) = (1 − tumor volume of aftertreatment/tumor volume of pretreatment) * 100%

Tumor Growth Curve

The tumor growth curve in different groups of the tumor-bearing mice onNCI-H1975, HCC827 and A431 models are shown in FIG. 6, FIG. 7, and FIG.8, respectively.

Tumor Weights

The mouse tumor weights in different groups on NCI-H1975, HCC827 andA431 models are shown in Table 17, Table 18, and Table 19, respectively.

TABLE 17 The Antitumor Activity of Compounds in the Treatment ofNCI-H1975 Model Tumor Weight (g) IR^(a) Treatment at day 23(14) at day23(14) p Vehicle 1.99 ± 0.16 — — Compound 3 25 mg/kg po qd 0.70 ± 0.0465.0% 0.001 Compound 3 50 mg/kg po qd 0.34 ± 0.08 82.8% 0.000 Gefitinib100 mg/kg po qd 1.69 ± 0.11 15.1% 0.717

TABLE 18 The Antitumor Activity of Compounds in the Treatment of HCC827Model IR^(a) Tumor Weight (g) at day Treatment at day 49(35) 49(35) pVehicle 1.94 ± 0.32 — — Compound 3 (PEG) 50 mg/kg po qd 0.06 ± 0.0296.7% .004 Compound 3 (MC) 50 mg/kg po qd 0.26 ± 0.04 86.5% .008Gefitinib 100 mg/kg po qd 0.03 ± 0.02 98.3% .004

TABLE 19 The Antitumor Activity of Compounds in the Treatment of A431Model Tumor Weight (g) IR^(a) Treatment at day 23(14) at day 23(14) pVehicle 1.67 ± 0.29 — — Compound 3 100 mg/kg po qd 1.60 ± 0.22  4.0%0.817 Gefitinib 100 mg/kg po qd 0.13 ± 0.03 92.5% 0.000 ^(a)IR(Inhibition Rate) = (TW_(Control) − TW_(Treatment))/TW_(Control) × 100%

Biological Example D Synthesis of the Maleate Salt and HydrochlorideSalt of Compound 3 and Pharmacokinetic Study

The synthesis of the maleate and hydrochloride salts from the free baseof Compound 3 is shown below:

To the free base of Compound 3 (2 g) in ethanol/water (5:95, 22 mL) at40° C., maleic acid (1.2 eq.) or HCl (2.2 eq.) was added dropwise. Afterthe solid was dissolved, the solution was cooled to room temperature,and stood for overnight. The resulting crystals (light yellow or offwhite) were collected, washed with cold water and dried overnight (over85% yield).

Pharmacokinetic Studies on Rats with Compound 3 in Free Base, MaleateSalt and HCl Salt Forms:

A pharmacokinetic comparison study was performed on rats using Compound3 in free base, maleate salt, and HCl salt forms. The detailed studyconditions along with the experimental results are shown the tablebelow:

TABLE 20 Cmax(po) Animal Dose AUClast or C₀ (iv) Compound FormulationSex Source (mg/kg) Route F % (ng/mL*h) t½ (h) (ng/ml) Compound 3PEG200:D5W Female Zhe Jiang 4.7 i.v. n/a  2195 ± 140.2  1.6 ± 0.044102.1 ± 675.8 Free base (50:50, v/v) AMS Compound 3 0.5% MC Female ZheJiang 30.56 p.o. 10.7 ± 2.6 1526.8 ± 364.6  2.4 ± 0.6 242.0 ± 37.6 Freebase AMS Compound 3 0.5% MC Female Zhe Jiang 40.88 p.o. 30.6 ± 8.25853.8 ± 1565.4 2.4 ± 1.0 1259.3 ± 359.0 HCl salt AMS Compound 3 0.5% MCFemale Zhe Jiang 42.00 p.o. 32.7 ± 9.8 6412.2 ± 1917.8 2.3 ± 0.4 1540.0± 528.5 Maleate salt AMS Compound 3 PEG200:D5W Female Zhe Jiang 37.6p.o.  23.0 ± 10.8 4041.0 ± 1892.3 2.8 ± 0.3 1263.3 ± 270.2 Free base(50:50, v/v) AMS

The results show that in the same formulation of 0.5% methylcellulose(MC), the salt forms (both maleate salt and HCl salt) had about a 3-foldbetter bioavailability compared to the free base form.

1-22. (canceled)
 23. A method of treating and/or preventing aproliferation disorder comprising administering to a subject in need ofsuch treatment an effective amount of at least one compound of Formula(VIII):

or a pharmaceutically acceptable salt thereof, wherein X¹ is O, NH, S,CH₂, or CF₂; R¹ and R² are independently selected from hydrogen, halo,C₁₋₆ alkyl, and C₁₋₆ haloalkyl; R³ is selected from halo, hydroxyl, C₁₋₆alkyl, C₁₋₆ alkoxy, cyano, and nitro; n is a number from zero to 4; R⁴is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;wherein the alkyl or cycloalkyl is unsubstituted or substituted withhydroxyl or amino; and wherein each R²² and R²³ are independentlyselected from hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined toform a 3 to 10 membered ring; R⁵ is selected from hydrogen and C₁₋₆alkyl; R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro; R⁷ is selectedfrom hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, hydroxyl, cyano, and nitro; R⁸ is selected from hydrogen,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,hydroxyl, cyano, and nitro; Q is CR⁹ or N; R⁹ is selected from hydrogen,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,hydroxyl, cyano, and nitro; R¹¹ is selected from hydrogen and C₁₋₆alkyl; R¹² is selected from hydrogen and C₁₋₆ alkyl; R¹³ is selectedfrom hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl, SO₂—C₁₋₆alkyl, C₃₋₇ cycloalkyl,and C₆₋₂₀ aryl; wherein each alkyl or aryl is unsubstituted orsubstituted with hydroxyl, C₁₋₆ alkoxy, or halo; and —NR¹⁸R¹⁹ is

wherein R¹⁰ is selected from hydrogen and C₁₋₆ alkyl; R¹⁵ isunsubstituted methyl, or is C₂₋₄alkyl unsubstituted or substituted withhydroxy, methoxy, or halo; and m is 1 or 2; or R¹⁹ and R⁹ taken togetherform a 5- or 6-membered heteroaryl ring optionally substituted withC₁₋₆alkyl that is unsubstituted or substituted with amino, hydroxyl, orhalo; and R¹⁸ is hydrogen or C₁₋₆alkyl, or is absent to satisfy valencyof the heteroaryl ring; provided that neither of R⁶ or R⁷ is methoxywhen —NR¹⁸R¹⁹ is


24. The method of claim 23, wherein the proliferation disorder isselected from the group consisting of sarcoma, epidermoid cancer,fibrosarcoma, cervical cancer, gastric carcinoma, skin cancer, leukemia,lymphoma, lung cancer, non-small cell lung cancer, colon cancer, CNScancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breastcancer, liver cancer, head and neck cancers, and pancreatic cancer. 25.The method of claim 23, further comprising administering an effectiveamount of a second prophylactic or therapeutic agent for treating and/orpreventing a proliferation disorder in a subject. 26-27. (canceled) 28.The method of claim 23, wherein the compound is a compound of Formula(Ia) or (Ib):

wherein R¹ and R² are independently selected from hydrogen, halo, C₁₋₆alkyl, and C₁₋₆ haloalkyl; R³ is selected from halo, hydroxyl, C₁₋₆alkyl, C₁₋₆ alkoxy, cyano, and nitro; n is a number from zero to 4; R⁴is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;wherein the alkyl or cycloalkyl is unsubstituted or substituted withhydroxyl or amino; and wherein each R²² and R²³ are independentlyselected from hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined toform a 3 to 10 membered ring; R⁵ is selected from hydrogen and C₁₋₆alkyl; R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro; R⁷ is selectedfrom hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkoxy, C₁₋₆haloalkoxy, hydroxyl, cyano, and nitro; R⁸ is selected from hydrogen,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,hydroxyl, cyano, and nitro; Q is CR⁹ or N; R⁹ is selected from hydrogen,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,hydroxyl, cyano, and nitro; R¹⁰ is selected from hydrogen and C₁₋₆alkyl; a is one or two; Ring A is an aromatic ring; R²⁰ and R²¹ areindependently selected from hydrogen and C₁₋₆ alkyl; wherein alkyl isunsubstituted or substituted with amino, hydroxyl, or halo; wherein R²¹may not present to satisfy valency; R¹¹ is selected from hydrogen andC₁₋₆ alkyl; R¹² is selected from hydrogen and C₁₋₆ alkyl; and R¹³ isselected from hydrogen, C₁₋₆ alkyl, C₁₋₆ acyl, SO₂—C₁₋₆alkyl, C₃₋₇cycloalkyl, and C₆₋₂₀ aryl, wherein each alkyl or aryl is unsubstitutedor substituted with hydroxyl, C₁₋₆ alkoxy, or halo; or apharmaceutically acceptable salt thereof.
 29. The method of claim 23,wherein the compound is a compound of Formula (II):

wherein R¹ and R² are independently selected from hydrogen, halo, C₁₋₆alkyl, and C₁₋₆ haloalkyl; R³ is selected from halo, hydroxyl, C₁₋₆alkyl, C₁₋₆ alkoxy, cyano, and nitro; n is a number from zero to 4; R⁴is selected from hydrogen, C₁₋₆ alkyl, C₃₋₇ cycloalkyl, and —NR²²R²³;wherein the alkyl or cycloalkyl is unsubstituted or substituted withhydroxyl or amino; and wherein each R²² and R²³ are independentlyselected from hydrogen and C₁₋₆ alkyl or R²² and R²³ may be joined toform a 3 to 10 membered ring; R⁵ is selected from hydrogen and C₁₋₆alkyl; R⁶ is selected from hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₂₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro; R⁷ is selectedfrom hydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkoxy, C₁₋₆haloalkoxy, hydroxyl, cyano, and nitro; R⁸ is selected from hydrogen,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,hydroxyl, cyano, and nitro; Q is CR⁹ or N; where R⁹ is selected fromhydrogen, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, hydroxyl, cyano, and nitro; R¹⁰ is selected from hydrogenand C₁₋₆ alkyl; R¹¹ is selected from hydrogen and C₁₋₆ alkyl; R¹² isselected from hydrogen and C₁₋₆ alkyl; and R¹³ is selected fromhydrogen, C₁₋₆ alkyl, and C₆₋₂₀ aryl, wherein each alkyl or aryl isunsubstituted or substituted with hydroxyl, C₁₋₆ alkoxy, or halo; or apharmaceutically acceptable salt thereof.
 30. The method of claim 23,wherein X¹ is O or NH.
 31. The method of claim 23, wherein R¹¹ ishydrogen.
 32. The method of claim 23, wherein R¹² is hydrogen.
 33. Themethod of claim 23, wherein R¹ and R² are each hydrogen.
 34. The methodof claim 23, wherein n is zero.
 35. The method of claim 23, wherein—NR¹⁸R¹⁹ is


36. The method of claim 23, wherein R¹⁰ is methyl.
 37. The method ofclaim 23, wherein R¹³ is hydrogen.
 38. The method of claim 23, whereinR¹³ is C₁₋₆alkyl substituted with hydroxyl or halo.
 39. The method ofclaim 23, wherein R⁶ is hydrogen.
 40. The method of claim 23, wherein R⁷is hydrogen or methoxy.
 41. The method of claim 23, wherein R⁸ ishydrogen.
 42. The method of claim 23, wherein Q is N.
 43. The method ofclaim 23, wherein Q is CR⁹.
 44. The method of claim 43, wherein R⁹ ishydrogen or fluoro.
 45. The method of claim 23, wherein R¹⁹ and R⁹together form a 5- or 6-membered ring optionally substituted withC₁₋₆alkyl that is unsubstituted or substituted with amino.
 46. Themethod of claim 23, wherein the compound selected from the groupconsisting of:

and pharmaceutically acceptable salts thereof.
 47. The method of claim23, wherein the compound is a compound of Formula (VII):

wherein R¹ and R² are independently selected from hydrogen, halo, C₁₋₆alkyl, and C₁₋₆ haloalkyl; R⁸ is selected from halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, hydroxyl, cyano, and nitro; R¹⁰is C₁₋₆ alkyl; and R¹³ is hydrogen or C₁₋₆ alkyl; or a pharmaceuticallyacceptable salt thereof.
 48. The method of claim 23, wherein thecompound is

or a pharmaceutically acceptable salt thereof.
 49. The method of claim48, wherein the compound isN-(3-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenyl)acrylamidehydrochloride salt or maleate salt.
 50. The method of claim 23, whereinthe compound inhibits a mutated EGFR in the subject.
 51. The method ofclaim 50, wherein the mutated EGFR comprises a T790M mutation.
 52. Themethod of claim 51, wherein the mutated EGFR comprises T790M/L858Rmutations.
 53. The method of claim 51, wherein the mutated EGFRcomprises delE746-A750 mutation.